Analysis of erythrocytic polyamines by automated reverse-phase liquid chromatography

✍ Scribed by Carl S. Killian; Farida P. Vargas; Gerald P. Murphy; Somnulu Beckley; T. Ming Chu

• Publisher: John Wiley and Sons
• Year: 1987
• Tongue: English
• Weight: 622 KB
• Volume: 1
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860010405

✦ Synopsis

A sensitive and reproducible method for quantitation of polyamines in erythrocytes by high-performance liquid chromatography (HPLC) is described. Spermidine and spermine are first converted to fluorescent dansyl derivatives and are then separated in less than 15 minutes on a C18 reversephase cartridge using methanol in water mobile phase. Sensitivity of the method is 20 pmol with recoveries that average 96% and 95% for spermidine and spermine, respectively. Precision study revealed an average mean coefficient of variation of 5.42 2.95 (mean f S.D.) from four levels of the polyamines (8.2 to 354.9 nmollg hemoglobin [Hgb]). The normal ranges for 30 apparently healthy men and women, aged 20-60 years, were 2.9-33.7 nmollg Hgb (f 2 S.D. from mean) for spermidine and 0-20.9 nmollg Hgb for spermine. The quantity of putrescine was negligible. Feasibility of this method was evaluated in serial specimens from an advanced-stage patient with prostate cancer who was receiving multiple-modality therapy. Results revealed that this HPLC method can be used in quantifying circulating polyamines in clinical specimens.