Beer–35

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TOTAL POLYPHENOLS
(International Method)
with closure).
Polyphenols in beer predominantly originate in (e) Magnetic stir plate.
malted barley and hops. They are water-soluble phenolic (f) Magnetic stir bar.
compounds that have the ability to precipitate alkaloids, (g) Volumetric flasks, 25-mL and 1,000-mL.
gelatin, and other proteins. There is evidence that (h) Graduated cylinders, 25-mL wide-neck, with glass
polyphenols are implicated in the complex phenomena stoppers.
of beer oxidation and haze formation. (i) Pipettes, 10-mL, 8-mL, and 0.5-mL.
The polyphenols are reacted with ferric ion in alkaline (j) Centrifuge, if needed for clarification of samples.
solution, and the red color produced is measured.
Method
Degas beer by shaking (see Beer Degassing Matrix).
Reagents
Clarify hazy worts or beers by centrifuging, not by
(a) Carboxymethylcellulose (CMC) sodium salt (low
filtration. For each sample analyzed, there will be a test
viscosity), CAS 9004-32-4.
and a blank 25-mL cylinder. These should be prepared
(b) Ethylenediamine tetraacetic acid (EDTA), CAS
at the same time. It is essential to mix thoroughly after
6381-92-6.
adding each reagent for both the test and the blank.
(c) Ferric ammonium citrate, CAS 1185-57-5. (d)
Pipet 10 mL of beer into both the test and blank
Ammonium hydroxide (28–30% as NH3), CAS
cylinders. Add 8 mL CMC/EDTA reagent (reagent 1)
1336-21-6.
into both the test and blank cylinders; cap with stopper
(e) Reagent water (18 MΩ resistivity or greater).
and mix. To the test cylinders only, add 0.5 mL ferric
ammonium citrate reagent (reagent 2), cap, and mix. To
Reagent Preparation
both the test and blank cylinders add 0.5 mL ammonium
(1) Carboxymethylcellulose (CMC/EDTA) reagent. A 1%
hydroxide solution (reagent 3), cap, and mix. Bring all
solution of sodium salt of CMC (low viscosity) with
cylinders to volume with reagent water and mix again.
the addition of 0.2% EDTA. In a large beaker
Allow solutions to stand for 10 min and measure
equipped with a magnetic stir bar, dissolve 2 g
absorbance in 10-mm cells at 600 nm against blank.
EDTA (reagent b) in approximately 800 mL reagent
Dilution of samples. When diluting sample for analy sis,
water.
use 50-mL graduated flask and, after adding
Mix on a stir plate and gradually add 10 g CMC
CMC/EDTA (reagent 1) and ferric ammonium citrate
(reagent a). When all solid material has dissolved
(reagent 2) reagents, add 25 mL distilled water, mix
(can take ~3 hr), transfer to 1-L volumetric flask
well, and follow with ammonium hydroxide solution
and make to volume with reagent water.
(reagent 3).
(2) Ferric ammonium citrate reagent. A fresh solution
of 3.5% green ammonium ferric citrate (16% Fe). Calculation
Dissolve 0.875 g ferric ammonium citrate (reagent c) Concentration of polyphenols in mg/L is given by
in ~15 mL of reagent water in a small beaker. A600 × 820 and is reported as a whole number.
Transfer to a 25-mL volumetric flask and dilute to
volume with reagent water. Should be prepared fresh Example
daily. Absorbance at 600 nm = 0.128.
(3) Ammonium hydroxide solution. Should be prepared
Polyphenols, mg/L
in a fume hood. In a 50-mL Erlenmeyer flask, mix
= 0.128 × 820
15 mL of ammonium hydroxide (reagent d) and 30 = 105
mL reagent water. Keep flask closed with stopper
until ready for use. Precision
In collaborative tests (1), repeatability coefficients of
Apparatus variation ranged from 2.17 to 3.28% and reproducibility
(a) Spectrophotometer, capable of operating at 600 nm. coefficients of variation ranged from 5.58 to 11.61%.
(b) Cuvettes, 10-mm.
(c) Beaker, large (1-L or larger) and small (~50-mL). (d) References
1. American Society of Brewing Chemists. Report of Sub
Erlenmeyer flask, 50-mL with stopper (or similar vessel committee on Polyphenols in Beer. Journal 36:129, 1978. 2.
European Brewery Convention. Analytica-EBC, 3rd ed. Method 7.7,
p. E64. Schweizer Brauerei-Rundschau: Zurich, Switzerland, 1975.

Released 1978, reviewed and revised 2015

doi: 10.1094/ASBCMOA-Beer-35