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

Journal of Chromatography. B, Biomedical Sciences and Applications
S FuX Liu

Abstract

A high-performance liquid chromatographic method for the determination of polyamines in human prostate has been developed. This method is based on pre-column derivatization with dansyl chloride (Dns-Cl). The derivatives were separated on a muBondapak C18 column (250x4.6 mm I.D.; 10 microm), and eluted with methanol and distilled water using a one-step linear gradient. The column eluate was monitored by fluorescence detection (excitation, 370 nm; emission, 506 nm). The within-assay precision of the study (C.V.) was as follows: putrescine (PUT) 2.88%, spermidine (SPD) 2.94% and spermine (SP) 1.17%. The between-assay precision (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 polyamine derivatives did not require extraction with an organic solvent such as ethanol, evaporation under vacuum or other condensation procedures. This is a simple, rapid and sensitive method that can be applied to the determination of polyamines in nearly all biological tissues and body fluids, such as urine and serum.

Citations

Jan 3, 2012·European Journal of Nutrition·Kang Uk YunSang Kyum Kim
Aug 14, 2001·Journal of Chromatography. B, Biomedical Sciences and Applications·P Campíns-FalcóL A Tortajada Genaro
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