Separation and Determination of Benzoylated Polyamines (Spermidine Andspermine) using HPLC Techniqe

Abstract

A simple and sensitive reversed-phase high performance liquid chromatography (Rp -HPLC) was developed for separation and determination of polyamines (Spermidine and Spermine). Derivatization of polyamines was improved by dissolving benzoyl chloride in methanol to increase the detection of UV-Visible detector at ? max. 254  nm. The benzoylated polyamines eluted by Rp- HPLC using methanol : water (52 : 48) as mobile phase. The detection limited of this method is (50ng/ml) .The separation was performed on BDS-DB (250 x 4.6 mm I.D) column at flow rate 1 ml/ min. using standard solutions. The retention time of two compounds (Spermidine and Spermine) were (7.7 , 8.5 min.) respectively, this method suggested that monitoring the Spermidine and Spermine compounds could play an important role in diagnosis of different types of carcinoma and fallow up chemo and radiotherapy treatments.