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 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.
Fu
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Central Laboratory, Affiliated Hospital of Shandong Medical University, Jinan, China.
Zou
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Wang
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Liu
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D002851:Chromatography, High Pressure Liquid; D003619:Dansyl Compounds; D006801:Humans; D008297:Male; D011467:Prostate / Q000737:chemistry*; D011700:Putrescine / Q000032:analysis*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013095:Spermidine / Q000032:analysis*; D013096:Spermine / Q000032:analysis*; D013696:Temperature; D013997:Time Factors
D003619:Dansyl Compounds; D013096:Spermine; C029038:dansyl chloride; D013095:Spermidine; D011700:Putrescine