2) Chromatography column for ion exchange, disposable
polyethylene.†
3) pH meter.
b. Reagents:
1) Organics-removal resin: Thoroughly rinse 16 to 50 mesh
resin‡ with deionized water and remove resin fines by decanting.
Rinse three times with pH 12 solution. Store resin in pH 12
solution and refrigerate to prevent bacterial growth.
2) Anion exchange resin: Add 100 to 200 mesh anion ex-
change resin* to a beaker and thoroughly rinse with deionized
water. Cover resin with 4NHCl, stir, and let settle. Decant and
repeat acid rinse twice more. Store resin in 4NHCl.
3) Hydrochloric acid, conc: Before use, bubble helium
through the acid for3hatrate of 100 mL/min. (CAUTION: Use
a fume hood.)
4) pH 1.6 solution: Adjust pH of deionized water to 1.6 with
HCl.
5) pH 12 solution: Adjust pH of deionized water to 12 with
KOH.
c. Procedure:
1) Organic removal—Place 5 cm washed resin in a 0.8-cm-ID
column. Precondition column, at 1 mL/min, with 30 mL pH 12
solution and 20 mL pH 1.6 solution. Using HCl and a pH meter
adjust sample to pH 1.6 to 1.8. Pass sample through preconditioned
column at rate of 1 mL/min. Discard first 10 mL and use next 11 to
50 mL collected for Se(IV) determinations by Methods 3114B or C,
or 3500-Se.C preceded, if Se(VI) also will be determined, by
3500-Se.B.5. If more than 50 mL sample are needed, use another
column or use a column with twice as much resin.
2) Iron removal—Place 4 cm prepared resin in a small chro-
matographic column (add resin to column filled with 4NHCl to
avoid air bubbles). Rinse column with 10 mL 4NHCl at flow rate
⬍6 mL/min. Let solution drain to top of resin, but do not let the
column run dry. Adjust sample to 4NHCl and pour into column.
Discard first 10 mL and collect the next 11 to 100 mL for Se(IV)
analysis by Methods 3114B or C, or 3500-Se.C preceded, if
necessary, by 3500-Se.B.2 and if Se(VI) also is to be determined,
by 3500-Se.B.5 below.
2.
Removal of Interference by Persulfate Digestion
The combination of this procedure with 3500-Se.B.5 and Meth-
ods 3114B or C, or 3500-Se.C is, in most cases, the preferred
method for determining total selenium in filtered water. A small
amount of ammonium or potassium persulfate is added to the
mixture of sample and HCl to remove interference from reducing
agents and to oxidize relatively labile organic selenium compounds
such as selenoamino acids and methaneseleninic acid.
If the sample contains hydrogen sulfide or a large concentration
of organic matter or is otherwise suspect or to confirm method
accuracy, reanalyze sample using digestion procedure 3 or 4 below.
Prepare 2% potassium or ammonium persulfate solution by
dissolving 2.0 g in 100 mL deionized water (prepare weekly).
Add 0.2 mL persulfate solution to the mixed sample and HCl of
3500-Se.B.5cbefore heating and proceeding with pretreatment
and analysis.
After completing analysis multiply concentration of selenium
determined in the acidified sample by 2.04 to obtain total sele-
nium in original sample.
3.
Removal of Interference by Alkaline Hydrogen Peroxide
Digestion
Occasionally, digestion with persulfate gives incomplete re-
covery of total selenium. In this case, digestion with hydrogen
peroxide is used to remove all reducing agents that might inter-
fere and to fully oxidize organic selenium to Se(VI). The result-
ing solution can be analyzed for total selenium after pretreatment
according to 3500-Se.B.5 below.
This method is suitable for determining total selenium in
unfiltered water samples, where particulate selenium is present.
When working with a new matrix, confirm results obtained by
reanalyzing the sample using 3500-Se.B.4.
a. Apparatus:
1) Beakers, 150-mL.
2) Watch glasses.
3) Hot plate.
4) Pipettor, 1-mL, and tips.
5) Graduated cylinder, 25-mL.
b. Reagents:
1) Hydrogen peroxide,H
2
O
2
, 30%. Keep refrigerated.
2) Sodium hydroxide, NaOH, 1N.
3) Hydrochloric acid, HCl, 1.5N: Dilute 125 mL conc HCl to
1 L with deionized water.
c. Procedure: Add 2 mL 30% H
2
O
2
and1mL1NNaOH to
25 mL sample in a beaker. Cover beaker to control spattering and
simmer on hot plate until fine bubbles characteristic of H
2
O
2
decomposition subside and are replaced by ordinary boiling. Add
1mL1.5NHCl to redissolve any precipitate that may have
formed, let cool, and pour into graduated cylinder. Rinse beaker
with deionized water into graduated cylinder and make volume
up to 25 mL. Proceed to 3500-Se.B.5 and chosen analytical
method.
4.
Removal of Interference by Permanganate Digestion
This digestion method utilizes potassium permanganate to
oxidize selenium and remove interfering organic compounds.
Excess KMnO
4
and MnO
2
are removed by reaction with hydrox-
ylamine. HCl digestion is included here, because it is conve-
niently performed in the same reaction vial. Selenite may then be
determined directly by Methods 3114B or C, or 3500-Se.C.
Verify recovery for the given matrix. This method gives good
recovery even with heavily contaminated water samples that
contain organic selenium compounds, dissolved organic matter,
and visible suspended material.
Permanganate may oxidize chloride ion to free chlorine. Part
of the chlorine (which interferes with hydride analysis) is re-
moved by reaction with hydroxylamine, but the best way to
eliminate free chlorine is by prolonged heating in an open vial
during the digestion step. Excess hydroxylamine may reduce
recovery by reducing selenium to Se(0).
a. Apparatus:
1) Oven with thermostat, for continuous operation at 110 ⫾
5°C.
2) Digestion vials, 40-mL, with fluorocarbon-lined screw caps.
† Bio-Rad Econo-Columns, or equivalent.
‡ Amberlite XAD-8, Supelco, or equivalent.
SELENIUM (3500-Se)/Sample Preparation
3
SELENIUM (3500-Se)/Sample Preparation