Liquid chromatographic determination of polyamines in human urine based on intramolecular excimer-forming fluorescence derivatization using 4-(1-pyrene)butanoyl chloride.
Liquid chromatographic determination of polyamines in human urine based on intramolecular excimer-forming fluorescence derivatization using 4-(1-pyrene)butanoyl chloride.
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基于 4-(1-芘)丁酰氯分子内准分子形成荧光衍生化的液相色谱测定人尿中的多胺。
DOI：
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
M. Yamaguchi
中科院分区：
文献类型：
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作者：
H. Yoshida;Hajime Harada;Y. Nakano;H. Nohta;J. Ishida;M. Yamaguchi
本文描述了一种用于高灵敏度、选择性荧光测定人尿中多胺（腐胺、尸胺、亚精胺和精胺）的液相色谱方法。该方法基于用芘试剂4 -（1 -芘基）丁酰氯（PBC）进行分子内形成激基缔合物的荧光衍生化，随后进行反相液相色谱分析。与先前报道的使用4 -（1 -芘基）丁酸N -羟基琥珀酰亚胺酯作为衍生化试剂的方法相比，此方法对亚精胺和精胺的测定具有更高的灵敏度。含有人尿稀释液中游离多胺的样品直接用PBC衍生化，并在辛基柱上分离。衍生物在激发波长345 nm和发射波长475 nm处检测。为测定总多胺含量，结合态多胺首先在4 M盐酸中水解。尿液中多胺的检测限（信噪比 = 3）为1.1 - 3.4 pmol/mL。在优化的衍生化和色谱条件下，生物源单胺等干扰物无峰出现，或者其峰不干扰多胺衍生物的峰。总之，本衍生化方法无需样品净化程序即可直接测定人尿样品中的多胺。
A liquid chromatographic method for highly sensitive and selective fluorometric determination of polyamines (putrescine, cadaverine, spermidine and spermine) in human urine is described. This method is based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(1-pyrene)butanoyl chloride (PBC), followed by reversed-phase liquid chromatography. The method offers higher sensitivity for determination of spermidine and spermine than previously reported method utilizing 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester as a derivatization reagent. Samples containing free polyamines in diluted human urine were directly derivatized with PBC and separated on an octyl column. The derivatives were detected at excitation 345 and emission 475 nm wavelengths. For determination of total polyamine content, the conjugated polyamines were first hydrolyzed in 4 M HCl. The detection limits (signal-to-noise ratio = 3) for polyamines in urine were 1.1-3.4 pmol/mL. At optimized derivatization and chromatographic conditions, interferences such as biogenic monoamines gave no peaks or the peaks did not interfere with the peaks of polyamine derivatives. In conclusion, the present derivatization method allows direct determination of polyamines in human urine samples without the need for sample clean-up procedures.
二芘基磷脂酰胆碱作为膜流动性探针。
影响因子：
3.4
作者：
Vauhkonen,M;Sassaroli,M;Somerharju,P;Eisinger,J
通讯作者：
Eisinger,J