采用高效液相色谱-荧光检测法测定人前列腺中的多胺。

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

作者信息

Fu S, Zou X, Wang X, Liu X

机构信息

Central Laboratory, Affiliated Hospital of Shandong Medical University, Jinan, China.

出版信息

J Chromatogr B Biomed Sci Appl. 1998 May 29;709(2):297-300. doi: 10.1016/s0378-4347(98)00068-1.

DOI:10.1016/s0378-4347(98)00068-1

PMID:9657227

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.

摘要

已开发出一种用于测定人前列腺中多胺的高效液相色谱法。该方法基于用丹磺酰氯（Dns-Cl）进行柱前衍生化。衍生物在μBondapak C18柱（内径250×4.6 mm；10μm）上分离，并用甲醇和蒸馏水通过一步线性梯度洗脱。柱洗脱液通过荧光检测（激发波长370 nm；发射波长506 nm）进行监测。该研究的批内精密度（变异系数）如下：腐胺（PUT）2.88%，亚精胺（SPD）2.94%，精胺（SP）1.17%。批间精密度（变异系数）为：PUT 2.66%，SPD 3.06%，SP 2.79%。回收率大于97%。PUT、SPD和SP的检测限分别为0.05、0.08和0.06 nmol/ml。与其他研究不同，样品或多胺衍生物不需要用乙醇等有机溶剂萃取、真空蒸发或其他浓缩程序。这是一种简单、快速且灵敏的方法，可应用于几乎所有生物组织和体液（如尿液和血清）中多胺的测定。