DETERMINATION OF POLYAMINES IN HUMAN PROSTATE BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH FLUORESCENCE DETECTION

A high-performance liquid chromatographic method for the determination of polyamines in human prostate has been developed. This method is ba sed on pre-column derivatization with dansyl chloride (Dns-Cl). The de rivatives were separated on a mu Bondapak C-18 column (250x4.6 mm I.D. ; 10 mu m), and eluted with methanol and distilled water using a one-s tep linear gradient. The column eluate was monitored by fluorescence d etection (excitation, 370 nm; emission, 506 nm). The within-assay prec ision of the study (C.V.) was as follows: putrescine (PUT) 2.88%, sper midine (SPD) 2.94% and spermine (SP) 1.17%. The between-assay precisio n (C.V.) was: PUT 2.66%, SPD 3.06%, SP 2.79%. The recovery was greater than 97%. The detection limit for PUT, SPD and SP were 0.05, 0.08 and 0.06 nmol/ml, respectively. In contrast to other studies, sample or p olyamine derivatives did not require extraction with an organic solven t such as ethanol, evaporation under vacuum or other condensation proc edures. This is a simple, rapid and sensitive method that can be appli ed to the determination of polyamines in nearly all biological tissues and body fluids, such as urine and serum. (C) 1998 Elsevier Science B .V. All rights reserved.