Measurement of acid-catalyzed isomerization of unsaturated aldehyde-2,4-dinitrophenylhydrazone derivatives by high-performance liquid chromatography analysis

Abstract

Aldehyde-2,4-dinitrophenylhydrazones exist as ( E)- and ( Z)-geometrical isomers, and adventitious isomerization during sample preparation can cause analytical errors. Purified alkenal-2,4-dinitrophenylhydrazone derivatives comprise only the ( E)-isomer. However, partial isomerization to the ( Z)-isomer occurs upon addition of acid to attain an equilibrium isomer ratio. The UV–visible spectral properties of the isomers differ; the ( Z)-isomer exhibiting a 6–10 nm lower absorption maximum compared to the ( E)-isomer. Alkenal-2,4-dinitrophenylhydrazones having a C C double bond at the 2- or 3-position of the alkenal exhibited similar absorption maxima with an equilibrium isomer ratio (0.035) that was much lower than those of other alkenals. The C C double bond at the 3-position migrates to a position of conjugation with the C N double bond during hydrazone synthesis to form a stabilized molecular structure. Alkenal-2,4-dinitrophenylhydrazones having a double bond at the 4-position or greater exhibited a similar absorption maxima equilibrium isomer ratio (0.14) to alkanal-2,4-dinitrophenylhydrazones. The quantitative analysis of carbonyl compounds in air or water using DNPH is usually conducted in the presence of an acid catalyst. Consequently, the solution of the direct extract prepared for HPLC or GC analysis contains both ( E)- and ( Z)-isomers.