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Separation by ion chromatography followed by conductivity detection.

Acetic acid (acetate) is converted to acetyl-CoA in the presence of the enzyme acetyl-CoA synthetase (ACS) by adenosine-5'-triphosphate (ATP) and coenzyme A (CoA).

\(\text{(1) Acetate + ATP + CoA }\space ^{\underrightarrow{\text{ACS}}} \space \space\text{Acetyl-CoA + AMP + Pyrophosphate}\)

Acetyl-CoA reacts with oxaloacetate in the presence of citrate synthase (CS) to form citrate.

\(\text{(2) Acetyl-CoA + Oxalacetate + H}_2\text{O }\space ^{\underrightarrow{\text{CS}}} \space \space\text{Citrate + CoA}\)

The oxaloacetic acid required for reaction (2) is produced from malic acid and nicotinamide adenine dinucleotide (NAD) in the presence of malate dehydrogenase (MDH). In doing so, NAD is reduced to NADH:

\(\text{(3) L-Malate + NAD}^+ \space ^{\underrightarrow{\text{L-MDH}}} \space \text{Oxalacetate + NADH + H}^+\)

The formation of NADH+H⁺ forms the basis of this analysis, which is measured as an increase in the absorbance at 340 nm. Since this concerns a previous indicator reaction, the quantity of NADH+H⁺ is not linearly proportional to the acetic acid concentration.

A cation exchanger based upon a sulfonated, crosslinked styrene/divinylbenzene copolymer is used to determine various organic acids in wort. Due to the high ligand density, the separation mechanism is based upon a combination of ion exclusion, ligand exclusion and steric exclusion; detection is performed using a UV detector.

Potassium dichromate oxidizes many organic and certain inorganic substances to various extents in an acidic solution. Since the level of oxidation depends upon the kinds of substances, the concentration of potassium dichromate, the pH of the solution, and the temperature and reaction time, the procedure described below must be followed precisely. The volume of potassium dichromate required in the analysis is determined potentiometrically. In an acidic solution, the dichromate ions are reduced to chromium(III) ions:

Cr₂O₇^2- + 6 e^- + 14 H₃O⁺ → 2 Cr³⁺ + 21 H₂O

Dichromate ions in excess of those required are determined through titration with an ammonium iron(II) sulfate solution:

Cr₂O₇^2- + 6 Fe²⁺ + 14 H₃O⁺ →2 Cr³⁺ + 6 Fe³⁺ + 21 H₂O 

Potassium permanganate oxidizes many organic and certain inorganic substances more or less completely in acidic, neutral or alkaline solutions. The volume of potassium permanganate required in the analysis is determined potentiometrically. Since oxidation depends on the type of solution, on its temperature and on the reaction time, the procedure described below must be followed precisely.

In acidic solutions, permanganate ions are typically reduced to manganese(II) ions:

MnO₄^- + 5 e- + 8 H₃O⁺ → Mn²⁺ + 12 H₂O

In alkaline solutions, the reduction results in tetravalent manganese only:

MnO₄^- + 3 e- + 4 H₃O⁺ → MnO₂ + 6 H₂O

Since in both cases the titration takes place in an acidic solution, this is irrelevant for the calculation. By adding oxalic acid, both the excess permanganate ions as well as the tetravalent manganese are reduced to manganese(II) ions:

2 MnO₄^- + 5 C₂O₄^2- + 16 H₃O⁺ → 2 Mn²⁺ + 24 H₂O + 10 CO₂

MnO₂ + C₂O₄²- + 4 H₃O⁺ → Mn²⁺ + 6 H₂O + 2 CO₂  

Total carbon (TC) and total inorganic carbon (TIC) are converted into carbon dioxide through oxidation (TC) or acidification (TIC). The CO₂ is transferred from the digestion cuvette through a membrane into the indicator cuvette. The color change of the indicator is measured in a photometer. TOC (total organic carbon) is determined as the difference between the values for TC and TIC.