Determination of the steam-volatile acids by means of titration
This method serves as a means for determining the titratable steam-volatile acids in beverages and concentrates
Volatile acids are distilled using steam, and the distillate is analyzed through titration. Sulfurous acid present in the distillate is determined iodometrically and subtracted from the total.
Determination of formic acid by enzymatic means
This analysis is suitable for wort, beer, beer-based beverages and NAB.
Formic acid can be found in beer in small quantities, and it is also formed by contaminating bacteria, for example, by lactic acid bacteria (rods).
In the presence of the enzyme formate dehydrogenase (FDH), the formic acid created in this reaction is quantitatively oxidized to bicarbonate by nicotinamide adenine dinucleotide (NAD):
HCOO- + NAD+ + H2O \(^{\underrightarrow{FDH}}\) HCO3- + NADH + H+
The amount of NADH formed during the reaction is equivalent to the amount of formic acid and can be determined photometrically based on its absorbance at 334, 340 or 365 nm.
Oxalic acid (oxalate) is transformed into formic acid and CO2 in this reaction catalyzed by the enzyme oxalate decarboxylase (Ox-DC):
Oxalic acid \(^{\underrightarrow{Ox-DC}}\) formic acid + CO2
The quantity of NADH produced during the reaction is equivalent to the amount of formic acid and is determined photometrically owing to its absorption at 334, 340 or 365 nm. In preparation for this test, the free formic acid present in the solution is determined along with the oxalic acid. Each are accounted for in the blank.