Determination of the total acidity through titration
This method is used to determine the total titratable acids in beverages and concentrates.
Titratable acidity represents the sum of the free acids present in a beverage, with the exception of the dissolved carbon dioxide (carbonic acid). In fruit juices and the beverages prepared from them, they usually consist of malic acid, citric acid and tartaric acid.
The titration of the degassed beverage sample (freed from carbonic acid) is carried out potentiometrically using 0.25 mol/l sodium hydroxide solution either to a pH of 7.0 calculated as tartaric acid or to a pH of 8.1 calculated as citric acid.
Determination of organic acids using ion chromatography
This method is suitable for beer, wort, green beer, NAB, water and wastewater
Separation by ion chromatography followed by conductivity detection.
The method describes how to determine the iso-α-acids, α-acids and β-acids in hop extracts and isomerized extracts by means of high-pressure liquid chromatography.
Hop extract and isomerized hop extract intended for use in beer brewing or elsewhere in the food industry
Hop extracts and isomerized hop extracts are dissolved in methanol. The iso-α-acids, α-acids and β-acids are separated using hig-pressure liquid chromatography (HPLC) with gradient elution and measured spectrophotometrically at a wavelength of 270 nm (iso-α-acids) and 314 nm (α-acids and β-acids).
This method describes how to determine iso-α-acids, α-acids and β-acids in isomerized pellets by means of reverse phase high pressure liquid chromatography (RP-HPLC).
Isomerized pellets intended for use in beer brewing or elsewhere in the food industry
The bitter substances in isomerized hop pellets contain a substantial amount of iso-α-acids; however, in addition to these, non-isomerized α-acids and β-acids are also present. In order to determine their content, a specific method is required.
After milling, the substances in question are extracted from the isomerized pellets using a diethyl ether/methanol mixture and a hydrochloric acid solution. The iso-α-acids, α-acids and β-acids dissolved in the ether phase are separated using reversed-phase high-performance liquid chromatography (RP-HPLC) and an elution gradient. They are then measured spectrophotometrically at wavelengths of 270 nm (iso-α-acids) and 314 nm (α-acids and β-acids).
Determination of oxalic acid by enzymatic means
Suitable for malt, wort, beer, beer-based beverages and soft drinks
Oxalic acid is primarily derived from malt. By reacting with the calcium ions in the brewing liquor, haze caused by calcium oxalate can form. These crystals also serve as nucleation sites for the spontaneous and rapid release of carbon dioxide (gushing). The precise determination of oxalic acid is therefore of great importance in brewing technology.
Oxalic acid (oxalate) is oxidized to carbon dioxide and hydrogen peroxide by the enzyme oxalate oxidase.
\(\text{ Oxalate} \hspace{0.5em}^{\underrightarrow{oxalatoxidase}}\hspace{0.5em} H_2O_2\hspace{0.3em}{+}\hspace{0.3em}CO_2\)
In the presence of the enzyme peroxidase (POD), hydrogen peroxide reacts with MTBH (3-methyl-2-benzo thiazolinone hydrazone) and DMAB (3-dimethyl amino benzoic acid to form a blue quinone complex.
\(H_2O_2+MTBH+DMAB\hspace{0.8em}^{\underrightarrow{POD}} \hspace{0.8em} \text{quinone complex} \space + \space H_2O\)
The intensity of the color is proportional to the concentration of the oxalate in the sample and is measured at 590 nm.
Determination of acetic acid by enzymatic means
This analysis is suitable for malt, wort, beer, beer-based beverages and soft drinks
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).
Acetate + ATP + CoA \(^{\underrightarrow{ACS}}\) Acetyl-CoA + AMP + pyrophosphate
Acetyl-CoA reacts with oxaloacetate in the presence of citrate synthase (CS) to form citrate.
Acetyl-CoA + oxaloacetate + H2O \(^{\underrightarrow{CS}}\) 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:
Malate + NAD+ \(^{\underleftrightarrow{L-MDH}}\) oxaloacetate + NADH + H+
The formation of NADH forms the basis of this analysis, which is measured as an increase in the absorbance at 340, 334 or 365 nm. Since this concerns a previous indicator reaction, the quantity of NADH is not linearly proportional to the acetic acid concentration.