Determination of polyamines in human saliva by high-performance liquid chromatography with fluorescence detection

A high-performance liquid chromatographic method for the determination of p olyamines (spermine, spermidine and putrescine) in human saliva was develop ed. This method is based on pre-column derivatization with o-phthaldialdehy de (OPA). The derivatives were separated on a Nucleosil ODS column (250x4.6 mm I.D.; 5 mum). The gradient elution was performed with two mobile phases A (water) and B (methanol) at a flow rate of 0.8 ml/min. The column eluate was monitored by fluorescence detection (excitation, 360 nm; emission, 510 nm). The within- and between-assay coefficients of variation for all the c ompounds were below 5%. The detection limits for spermine, spermidine and p utrescine were 0.04, 0.05 and 0.06 nmol/ml, respectively. The recovery was greater than 90%. Our analytical technique requires neither preliminary ext raction with an organic solvent, nor long multi-step procedures. For saliva samples, this is a simple, rapid and highly reproducible method that can b e easily applied to the routine determination of salivary polyamines, whose levels increase early in several pathological conditions. (C) 2001 Elsevie r Science B.V. All rights reserved.