Analytical Method: Selenium in water

Instuments and Reagents:

Cat.No. 109748  Spectroquant® Photometer NOVA 30 or

Cat.No. 109751  Spectroquant® Photometer NOVA 60

Cat.No. 114944  Rectangular cells 50 mm
Cat. No. 100264 Formic acid 98-100% for analysis
3,3‘-Diaminobenzidine tetrahydrochloride (CAS 868272-85-9), e. g. Sigma-Aldrich Art. 32750
Cat.No. 109992  TitriplexIII® solution 0.1 mol/l 
Cat.No. 105432  Ammonia solution 25% for analysis
Cat.No. 108389  Toluene for gas chromatography
Cat.No. 109584  MColorpHastTM pH-indicator strips, pH 2.0 - 9.0
Cat.No. 106649  Sodium sulfate anhydrous for analysis
Cat.No. 116754  Water for analysis
Membrane filter with 0.45 μm pore diameter


Measuring range:
0.05 – 2.00 mg/l
Reagent 1

Fill 9.4 ml formic acid 100% in a calibrated or conformity-checked 100-ml volumetric flask. Make up to volume with water for analysis and mix.


Reagent 2

Dissolve 0.5 g 3,3’-diaminobenzidine tetrahydrochloride in 100 ml chilled water for analysis. Prepare freshly each day and store in an amber glass bottle.


Reagent 3

Fill 48 ml ammonia solution 25% for analysis in a calibrated or conformity-checked 100-ml volumetric flask. Make up to volume with water for analysis and mix.


Sample preparation:
The sample must be clear; if not, filter through a membrane filter with 0.45 μm pore diameter.


Mix 30 ml nearly neutral sample solution with 2 ml reagent 1, 2 ml Titriplex III® solution 0.1 mol/l and 2 ml reagent 2. Adjust the pH value of the mixture with reagent 3 to 2.5 and set aside for 45 min at a dark place. Adjust the pH value of the mixture again with reagent 3 to 7.0, transfer into a 100-ml separation funnel, add 15 ml water for analysis and 7 ml toluene for analysis. Shake the mixture in the closed funnel for 1 min. Allow to separate and discharge the lower water phase. Fill the toluene extract in a test tube, add some anhydrous sodium sulfate and shake. Fill the clear, dried toluene extract in a 50 mm rectangular cell and measure at 445 nm against pure toluene.


Parameter list:

Method number


Method designation



445 nm







Zero point (E0)


Slope (E/C)


Meas. rang begin


Meas. range end


Reference cell

50 mm rect. cell

Calculation when measuring the absorbance value using NOVA photometers without programming the method in the memory of the instrument:

Concentration [mg/l] = Absorbance x 0.654 – 0.037

Result = measured value in mg/l Se4+

Statistical Data:

Standard deviation: 0.019 mg/l

Coefficient of variation: 2.9 %


Reference: J. Fries, H. Getrost.Organische Reagenzien für die Spurenanalyse. Merck 1975.  



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