CALCIUM DETERMINATION IN WORT

& BEER

1. Summary.
This instruction describes the titrimetric method for determination of calcium in wort and
beer. The concentration of calcium ion is expressed in mg/l.

2. Reference.

3. Definition.

4. Background.
4.1. Calcium in wort has a function to settle yeast in fermenting process and precipitate the
oxalate in beer, which form sediment in the finished product. 50 – 60 mg/l of Calcium in
cold wort is an optimum concentration.
4.2. Calcium is determined through the titration with Titriplex solution B (Na 2EDTA) and
Eriochrome black T as an indicator at pH 10.3. Color of solution changes from purple to
dark green at the endpoint.
4.3. At pH = 10.3, both of Calcium and Magnesium ions bind Na 2EDTA, so the determination
has to carry on two steps separately :
- Determination of total Ca and Mg by Titriplex solution at pH = 10.3.
- Separation of Calcium in wort by precipitating oxalate in the fridge for two hours, and
determination of Magnesium remains in the solution by Titriplex solution at pH = 10.3.
- The amount of Calcium in sample is a subtraction of Mg from total.

5. Applicability.
This method is applicable for wort & beer.

6. Equipment.
6.1. Refrigerator.
6.2. Measuring cylinder, 100 ml.
6.3. Flask, 250 ml.
6.4. Bulb pipette, 20.00 ml.
6.5. Funnel and pleated filter paper (fine paper).
6.6. Burette.

7. Reagents.
7.1. Titriplex solution C: 0.01 M
7.2. Eriochrome black T indicator.
Mix 100 mg Eriochrome black T with 30 g potassium chloride in a mortar. Transfer to
storage container.
7.3 Buffer solution pH = 10.3.
Dissolve 135 g ammonium chloride in 1.5 L distilled water. Add 875 ml 25 % ammonia
solution. Mix and measure the pH. Correct pH = 10.3 by 1.0 N NaOH or 1.0 N HCl
solution.
7.4 Magnesium-EDTA solution 0.3 %.
Dissolve 3 g MgEDTA with the distilled water in a 1000 ml measuring flask. Fill up to
the mark and mix. Transfer to storage Buffer solution pH = 10.3.
7.5 Saturated Ammonium Oxalate solution.
 CALCIUM DETERMINATION IN WORT
& BEER

Dissolve 10 gr of ammonium oxalate in 100 ml of distilled water. Shake well until the
crystals can not dissolve any more. After the crystals settle, decant the liquid into the
flask with label for use.
7.6 Hydrochloric acid 4N. For preparation,.

8. Procedure.
8.1. Sample 1
8.1.1. Take 50 ml of wort into a 250 ml flask.
8.1.2. Add 1.0 ml saturated ammonium oxalate solution and 4 drops buffer pH=10.3, and mix
well.
8.1.3. Cover flask by Para film and put it in a refrigerator for 2 hours.
8.1.4. Filter this sample through the pleated filter paper without filter powder. Filtrate should be
clear, if not circulate filtrate several times.
8.1.5. Pipette 20 ml of clear filtrate into a 250-ml flask and then add 4 drops of 4N HCl solution,
then add 80 ml of distilled water and mix (flask 1)
8.2. Sample 2
8.2.1. Filter wort through the pleated filter paper without filter powder.
8.2.2. Pipette 20 ml of clear filtrate into a 250-ml flask.
8.2.3. Add 80 ml of distilled water and mix (flask 2).
8.3. Add 10 ml buffer pH = 10.3 and 5 ml MgEDTA solution 0.3 % into flask 1 and flask 2.
8.4. About approx. 70 mg of Eriochrome black T indicator is added into each flask and titrates
with Titriplex C solution until color changes from purple to dark green.
Note: The indicator should be added just before titration.
8.5. Write down the volume of Titriplex solution C used.

9. Calculation.
9.1. The concentration of Calcium ions in wort

Ca 2+ conc = (V2-V1)*19.992 (mg/l)

V1: volume of Titriplex used for flask 1

V2: volume of Titriplex used for flask 2

10. Reporting.
10.1. Record the result(s) in mg/l in the Qualass software and / or in the report sheet.
10.2. Rounding off to one decimal place.

11. Safety & Remarks.

11.1. Concentrated hydrochloric acid is a strong acid that gives off a fume. It may only be used in
the fumigation chamber.
11.2. Drain out several ml of Titriplex before titrating to make sure that there are no bubbles in
the tube.
11.3. Temperature of refrigerator should be from 5 to 8°C, if not the precipitation of calcium
oxalate is uncompleted.
CALCIUM DETERMINATION IN WORT
& BEER