B-590.35.112 [2020-10] Oxalic Acid (Oxalate) – Enzymatic Method

The positive effects of fermented beverages on human health have been known for hundreds for years. For this reason, the cult drinks kvass (Russia) and kombucha (Asia) can be traced over a long history and have been consumed for their healing powers through time. Microorganisms such as lactic acid and acetic acid bacteria are used for non-alcoholic fermentation. The products of fermentation include organic acids such as lactic acid and gluconic acid, which facilitate digestion and metabolic processes. Fermented beverages are especially favored by consumers as healthy, natural refreshments due to their slightly sour flavor.

In addition to other criteria, the acid spectrum typical for certain types of fruit serves as the tool for evaluating whether a fruit juice is pure, and its composition has not been altered. Generally, tartaric acid, citric acid and L-malic acid are analyzed. These are the primary acids, with a few exceptions, that are responsible for the total acid in fruits.

Application/Purpose

Determination of oxalic acid by enzymatic means

Scope of Application

Suitable for malt, wort, beer, beer-based beverages and soft drinks

Principle

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.

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