Fluorometric Enzymatic Method for Determination of Formaldehyde

Abstract

Two sensitive fluorometric enzymatic methods for the determination of formaldehyde are described. These methods were developed using the enzyme formaldehyde dehydrogenase to catalyze the oxidation of formaldehyde to form formate and NADH in the presence of β-nicotinamide adenine dinucleotide (NAD⁺). In method I, the increase in NADH, which is directly proportional to the concentration of formaldehyde, is measured fluorometrically at λex= 348 nm and λem= 467 nm. In method II, the NADH is subsequently reacted with resazurin in the presence of a second enzyme diaphorase to form resorufin, a highly fluorogenic compound. The fluorescence intensity is then measured at λex= 575 nm and λem= 590 nm. The optimum conditions as well as the linear range and precision of these methods were investigated.

Key Words:

• Fluorometric Method
• Formaldehyde
• Formaldehyde Dehydrogenase