9.2.

41
AOAC Official Method 993.23
Dissolved Hexavalent Chromium
in Drinking Water, Ground Water,
and Industrial Wastewater Effluents
Ion Chromatographic Method
First Action 1993
(Applicable to determination of 1.2960g/Lhexavalent chromium
in drinking water, ground water, and industrial wastewaters.)
Caution: Hexavalent chromium is toxic and possibly carcino-
genic.
Results of interlaboratory study:
See Table 993.23 for interlaboratory study data.
A. Principle
After pH of filtered test sample is adjusted to 99.5, Cr(VI), as
CrO
4
2
, is separated by ion chromatography (IC) using anion ex-
change separat or col umn. Cr(VI) i s deri vat i zed wi t h
diphenylcarbazide in post-column reactor and detected as colored
complex at 530 nm.
B. Apparatus
(a) ICsystem.Equipped with pump capable of 2000 psi at con-
stant flow of 15 mL/min and containing no metal parts in sample,
eluant, or reagent flow path. Plastic pressurized eluant container,
12 L. Various sample loops, 50250L. Pressurized post-column
derivatization reagent delivery module with mixing tee and beaded
mixing coil capable of constant flow, 0.5 mL/min. Detector with
low-volume flow-through cell containing no metal parts in contact
with eluant flow path, capable of 530 nm. Recorder, integrator, or
computer to receive analog or digital signals for detector response
(peak height or area) as function of time. Chromatographic condi-
tions: eluant flow rate, 1.5 mL/min; post-column flow rate,
0.5 mL/min; typical Cr(VI) retention time, 3.8 min.
(b) Columns.(1) Guard column.For use with wastewater
materials. Capable of removing strongly adsorbing organics and
particles that would damage separator column (Dionex IonPac NG1,
Dionex Corp., Sunnyvale, CA, is suitable). (2) Separator col-
umn.Packed with high-capacity anion exchange resin capable of
separating CrO
4
2
from other sample constituents (Dionex IonPac
AS7 is suitable).
(c) Glassware.Class A, volumetric flasks, graduated cylinder,
and calibrated pipettes.
(d) Syringes.10 mL, male Luer-lock, disposable.
(e) Syringe cartridge filters.0.45 m, 7.3 cm diameter,
polysulfone filter (Gelman Acro 50A is suitable).
(f) Storage bottle.1 L, high-density polypropylene.
(g) Sample bottle.125 mL, high-density polypropylene.
(h) pH meter.Capable of determining pH to accuracy of
0.03 unit.
C. Reagents
(a) Ammonium hydroxide (NH
4
OH).
(b) Ammonium sulfate [(NH
4
)
2
SO
4
].
(c) 1,5-Diphenylcarbazide (DPC).
(d) Methanol (CH
3
OH).HPLC grade or equivalent.
(e) Sulfuric acid (H
2
SO
4
).Concentrated (specific gravity 1.84).
(f) Reagent water.ASTM Type I water (ASTM D1193). Suit-
able water may be obtained by passing distilled water through mixed
bed of anion and cation exchange resins.
(g) Sodium chromate tetrahydrate (Na
2
CrO
4
4H
2
O).
(h) Stock solution Cr(VI).Accurately weigh 4.501 g Na
2
CrO
4

4H
2
O, (g), to nearest 0.001 g, dissolve, and dilute to volume with re-
agent water, (f), in 1 L volumetric flask. Transfer to polypropylene
storage bottle, B(f).
(i) Eluant.250 mM (NH
4
)
2
SO
4
, 100 mM NH
4
OH. Dissolve ca
33 g (NH
4
)
2
SO
4
, (b), in 500 mL reagent water, (f); add 6.5 mL
NH
4
OH, (a); and dilute to volume with reagent water in 1 Lvolumet-
ric flask.
(j) Post-column reagent.2 mMDPC, 10%(v/v) CH
3
OH, .05M
H
2
SO
4
. Dissolve ca 0.5 g 1,5-diphenylcarbazide, (c), in 100 mL
CH
3
OH, (d). Add to 500 mL reagent water, (f), containing 28 mL
concentrated sulfuric acid, with stirring. Dilute to volume with re-
agent water in 1 L volumetric flask. Stable 45 days. Prepare as
needed.
(k) Buffer solution.Dissolve ca 33 g (NH
4
)
2
SO
4
, (b), in 75 mL
reagent water, (f), and add 6.5 mL NH
4
OH, (a). Dilute to 100 mL
with reagent water.
D. Material Collection, Preservation, and Storage
Filter test sample through 0.45mfilter, B(e). Wet syringe filtra-
tion unit using portion of test sample, then filter and collect required
volume of filtrate test portion 1. Add buffer, C(k), dropwise, to ad-
just test portion pH to 9.09.5, periodically checking pH with pH
meter. Ca 10 mL test portion is sufficient for 3 analyses.
Ship and store laboratory samples at 4C. Warm to ambient tem-
perature prior to analysis. Analyze laboratory samples within 24 h of
collection.
E. Calibration
Establish IC operating conditions, B(a). Prepare calibration stan-
dards at 3 concentration levels, bracketing anticipated concentration
range of samples. Dilute aliquots of stock standard, C(h), with re-
agent water, C(f), in volumetric flasks. Adjust calibration solutions
to pH9.09.5 with buffer solution, C(k), prior to final dilution. Pre-
pare low-level calibration standards daily.
To construct calibration curve, plot analyte response (peak height
or area) vs analyte concentration. Do not prepare calibration curve
over range >2 orders of magnitude.
Verify continuing calibration, using laboratory blank and calibra-
tion check standard (mid-level concentration) as surrogate samples
every 10 analyses. If concentration of analyte deviates from true
concentration by greater than 5%, recalibrate and reanalyze previ-
ous 10 samples.
Verify calibration stock standards, C(h), every 3 months by ana-
lyzing standard prepared fromUSEPA(U.S. Environmental Protec-
tion Agency) Certified Reference Material, or equivalent, using
acceptance limits of 10% of established value. If test portion re-
sults are outside acceptance limits, perform second analysis. If sec-
ond value exceeds acceptance limits, stop analyses, find source of
problem, and recalibrate instrument.
F. Procedure
(a) Equilibrate stored laboratory samples to ambient temperature
prior to analysis.
(b) Initiate instrument operating configuration and calibrate.
2000 AOAC INTERNATIONAL
(c) Drawca 3 mLtest portion into new, unused syringe, B(d), and
attach syringe filter, B(e). Discard first 0.5 mL through filter and
load 10material loop volume to thoroughly flush loop. If concen-
trations are higher than established linear dynamic range, dilute test
portions using reagent water, C(f), and adjust pH if necessary.
G. Calculations
Determine peak height or peak area counts of Cr(VI) in each test
portion. Compare with calibration curve to determine sample con-
centration in g/L.
H. Quality Control
Minimum QC requirements:
(1) To determine method detection limit (MDL), spike 7 replicate
portions of reagent water, C(f), at concentration 25estimated de-
tection limit of Cr(VI). Process replicate portions through entire ana-
lytical method, F, then analyze. Calculate MDL as follows:
MDL = t s
where t = Students t value, 99% confidence level, n - 1 degrees of
freedom(t =3.143 for 7 replicates) and s =standard deviation of rep-
licate analyses. MDLis concentration that can be distinguished from
background by analytical instrumentation used.
(2) Analyze method blank daily for contamination.
(3) Analyze 1 laboratory spiked water blank at 40g/L for each
10. Recovery from test portion must be within 3644g/L, or cor-
rective action should be taken and documented before analyses con-
tinue.
(4) Analyze 1 spiked matrix portion for each 10 portions. Spike
concentration should be 40 g/L or 2 background concentration,
whichever is higher. Calculate percent recovery of analyte spike. If
results are not within 90100%, reanalyze spiked and unspiked test
portions to document matrix effect.
Reference: J. AOAC Int. 77, 994(1994).
2000 AOAC INTERNATIONAL
Table 993.23 Interlaboratory study results for determination of dissolved hexavalent chromium in drinking water, ground water,
and industrial wastewater effluents by ion chromatographic method
a
Spike, g/L Mean recovery, g/L s
r
s
R
RSD
r
, % RSD
R
, %
Reagent water
1.2
1.6
1.50
1.87 0.15 0.57 9.0 33.8
6
8
6.68
8.64 0.53 1.06 6.9 13.9
16
20
17.4
21.4 0.77 2.28 4.0 11.8
100
140
101
143 3.8 4.3 3.1 3.5
800
960
818
966 12.7 21.2 1.4 2.4
Other water
b
6
8
5.63
7.31 0.55 1.20 8.2 17.8
16
20
15.1
19.8 1.85 1.85 10.4 10.4
100
140
98.9
138 3.3 4.8 2.8 4.0
800
960
796
944 27.1 66.6 3.1 7.6
a
Samples analyzed as Youden pairs.
b
This includes drinking water, ground water, and industrial wastewaters.