Abstract
Summary
6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC), a recently discovered derivatizing reagent for amines, is presented to analyze biogenic amines, in particular polyamines using reversed-phase high performance liquid chromatography (HPLC). Adducts of AQC and polyamines were characterized regarding their spectroscopic properties and their suitability for fluorescence detection. ...
Abstract
Summary
6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC), a recently discovered derivatizing reagent for amines, is presented to analyze biogenic amines, in particular polyamines using reversed-phase high performance liquid chromatography (HPLC). Adducts of AQC and polyamines were characterized regarding their spectroscopic properties and their suitability for fluorescence detection. Conditions for a derivatization of amines with AQC are discussed in detail and possible fluorescent by-products within the derivatization process are described. Further, extraction procedures for amines and polyamines from various tissues and matrices prior to AQC derivatization are presented and stability of N-acetylated polyamines within acidic precipitation was investigated. An overview of HPLC methods for the analysis of polyamines and other amines is given with their respective data of validation. Additional, recovery study of polyamines from various tissues is discussed and further, potential interferences of adducts of AQC with commonly used inhibitors of polyamine metabolism were presented. AQC is a powerful derivatization reagent for amines forming stable and highly fluorescent derivatives. The reagent reacts with primary as well as secondary amines under mild alkaline conditions within a broad pH range. The presented HPLC methods for a chromatographic separation of AQC adducts with polyamines demonstrated high accuracy as well as reproducibility and is suitable for analysis of unconjugated and acetylated polyamines without interference of fluorescent byproduct.
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