SOLID-PHASE EXTRACTION AND DETERMINATION OF DANSYL DERIVATIVES OF UNCONJUGATED AND ACETYLATED POLYAMINES BY REVERSED-PHASE LIQUID-CHROMATOGRAPHY - IMPROVED SEPARATION SYSTEMS FOR POLYAMINES IN CEREBROSPINAL-FLUID, URINE AND TISSUE
SOLID-PHASE EXTRACTION AND DETERMINATION OF DANSYL DERIVATIVES OF UNCONJUGATED AND ACETYLATED POLYAMINES BY REVERSED-PHASE LIQUID-CHROMATOGRAPHY - IMPROVED SEPARATION SYSTEMS FOR POLYAMINES IN CEREBROSPINAL-FLUID, URINE AND TISSUE
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DOI：
10.1016/s0378-4347(00)83621-x
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发表时间：
1986-07-11
期刊：
影响因子：
--
通讯作者：
MARTON, LJ
中科院分区：
文献类型：
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作者：
KABRA, PM;LEE, HK;MARTON, LJ
本文介绍了一种灵敏、简便的液相色谱荧光检测法测定生物体液中的非共轭多胺和乙酰化多胺。用丹磺酰氯柱前衍生后，通过Bond-Elut C18柱洗脱提取未缀合的多胺和乙酰化多胺，然后在反相柱上梯度洗脱分离。未结合腐胺，亚精胺和精胺在水解尿液，脑脊液或组织的完整分析可以在20-26分钟内完成，而未结合的多胺和单乙酰多胺的同时分析可以在40分钟内完成。未水解的尿液和脑脊液需要一个键合洗脱阳离子交换净化丹磺酰化前。测定的标准曲线线性高达20 nmol/ml，日内和日间变异系数分别为1.1和4.6%之间和1.6和11.8%之间，分别。将该方法获得的结果与尿液、组织和脑脊液样本的成熟改良氨基酸分析仪方法获得的结果进行比较。两种方法的相关系数在0.933 ~ 0.996之间。非共轭和乙酰化多胺的检测限在50和150 fmol之间。在20多种可能干扰测定的药物和胺中，仅发现去甲间苯二甲胺与内标物1，6-二氨基己烷具有相同的保留时间。
A sensitive and simple liquid chromatographic assay with fluorometric detection for unconjugated and acetylated polyamines in biological fluids is described. After precolumn derivatization with dansyl chloride, unconjugated polyamines and acetylated polyamines were extracted by elution from a Bond-Elut C18 column and then separated on a reversed-phase column with gradient elution. The complete analysis of unconjugated putrescine, spermidine and spermine in either hydrolyzed urine, cerebrospinal fluid or tissue could be accomplished within 20-26 min, while the simultaneous analysis of unconjugated polyamines and monoacetylpolyamines could be completed within 40 min. Unhydrolyzed urine and cerebrospinal fluid required a Bond-Elut cation-exchange clean-up before dansylation. Standard curves for the assay were linear up to 20 nmol/ml, and the within-day and day-to-day coefficients of variation were between 1.1 and 4.6% and between 1.6 and 11.8%, respectively. Results obtained with the method were compared with results obtained with a well established modified amino acid analyzer method for urine, tissue and cerebrospinal fluid samples. The correlation coefficients between these two methods were in the range 0.933-0.996. Detection limits between 50 and 150 fmol were achieved for unconjugated and acetylated polyamines. Of more than twenty drugs and amines tested for possibly interference with the assay, only normetanephrine was found to have the same retention time as the internal standard 1,6-diaminohexane.