Measuring phosphate with the colorimeter

Vanadate-molybdate method

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Vanadate-molybdate method

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This method is based on APHA Standard Method 4500-P C. Description of the assay from the method: In a dilute orthophosphate solution, ammonium molybdate reacts under acid conditions to form a heteropoly acid, molybdophosphoric acid. In the presence of vanadium, yellow vanadomolybdophosphoric acid is formed. The intensity of the yellow color is proportional to phosphate concentration.

Reagents & Equipment

  • Colorimeter
  • Cuvettes
  • Vanadate-molybdate reagent, Labchem Cat # LC266002
  • 1 mL variable micropipette

Colorimeter set-up

  • Wavelength: 470 nm. Use BLUE led (colorimeter RGB board).
  • Phosphate standard data: download a data file or prepare your own phosphate standard data
  • Before measuring samples, calibrate the colorimeter against water or a distilled water sample developed with vanadate-molybdate reagent as described below

Method

  • Transfer 1 mL of sample water into a clean cuvette.
  • Add 0.25 mL of vanadate-molybdate reagent and mix by pipetting up and down several times.
  • Wait at least 10 minutes for color to develop.
  • Place the cuvette with your sample into the colorimeter and click measure. The program will return the phosphate concentration in ppm.

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