The use of Girard-T reagent in a rapid and sensitive method for measuring glyoxal and certain other α-dicarbonyl compounds

Abstract

Girard-T reagent (trimethylaminoacetohydrazide chloride) under mild acid or alkaline conditions, reacts with aldehydes and ketones containing α-dicarbonyl functional groups to produce addition compounds which strongly absorb ultraviolet light. The glyoxal adduct has a λ max of 295 nm and an E max of 2.73 × 10 4 at pH < 6; it has a λ max of 325 nm and an E max of 1.88 × 10 4 at pH > 9. As little as 5 nmol of glyoxal can be measured accurately; absorbance is linear with concentrations up to at least 3500 nmol, and the reaction is complete within 10 min. A variation of the method is described which permits the detection of certain α-dicarbonyl compounds on paper. Another variation allows the measurement of glyoxal in a glyoxal-guanylic acid adduct, a previously described modified nucleotide.