An Improved High Performance Liquid Chromatographic Method for Identification and Quantization of Polyamines as Benzoylated Derivatives

Abstract

A simple reversed phase high performance liquid chromatography (RP-HPLC) method was developed for the determination of putrescine, cadaverine and spermidine (a class of polyamines) in their benzoylated form from external known standards. In the optimization procedure, a number of parameters were examined: 1) Solvent used in the extraction of standard polyamines (diethyl ether versus chloroform); 2) Solvent used in the elution of the polyamine (methanol versus acetonitrile); 3) Mode of derivatization and extraction step(s) (derivatization and extraction performed together versus derivatization and extraction performed separately); and 4) Other instrumental parameters (such as UV detection wavelength, gradient profiles). The advantages of our method, relative to the standard Morgan method are: a) decreased chro-matographic runtime, b) ease of preparation with good resolution, sensitivity, and reproducibility using a standard RP-HPLC method.