Analysis of Polyamines and Their Acetylated Forms with 9-Fluorenylmethyl Chloroformate and Reversed Phase HPLC

Abstract

A simple, fast and sensitive reversed phase HPLC method has been developed for analysis and quantitation of acetylated and non acetylated forms of polyamines. the polyamines are pre-column derivatized with 9-fluorenylmethyl chloroformate (FMOC) and detected in fluorescence. the derivatization reaction is completed in less than two minutes. the excess of FMOC is derivatized with glycine which reduces the interference from the non reacted reagent to a minimum. the conditions favor separation and detection of N¹-acetylputrescine, putrescine, N⁸-acetylspermidine and N¹-acetylspermine in a standard of 10 fmol. Spermidine and spermine are detected at 100 amol. Here, the method is used for analysis of extracellular polyamines in synovial fluid from patients with rheumatoid arthritis and for intracellular detection of rabbit lens polyamines.