Kilts C D, Gooch M D, Knopes K D
J Neurosci Methods. 1984 Sep;11(4):257-73. doi: 10.1016/0165-0270(84)90088-8.
A sensitive and specific procedure for quantifying plasma catecholamines using on-line trace enrichment high performance liquid chromatography with electrochemical detection (HPLC-EC) is described. Plasma catecholamines are off-line extracted using aluminum oxide and then on-line enriched on cation exchange enrichment HPLC columns and back-eluted using a potassium enriched mobile phase for further separation by ion-pair, reverse phase HPLC. The salient features of this method are (1) the on-line combination of distinct HPLC retention mechanisms for enrichment and separation of the catecholamines to yield optimal assay specificity, (2) a lower working limit of detection of 25 pg norepinephrine and epinephrine per milliliter plasma based upon an assayed plasma volume of 1 ml, (3) the generation of calibration curves using standard additions to a 'like' matrix, (4) a high daily sample throughput due to the lack of late-eluting chromatographic interferences and (5) use of an optimized aluminum oxide prepurification procedure. The advantages of this method relative to other HPLC-EC assays for plasma norepinephrine and epinephrine are discussed as are the general considerations to be addressed when defining performance characteristics of an assay.
本文描述了一种灵敏且特异的方法，用于使用在线痕量富集高效液相色谱-电化学检测法（HPLC-EC）对血浆儿茶酚胺进行定量分析。血浆儿茶酚胺首先使用氧化铝进行离线提取，然后在线富集于阳离子交换富集HPLC柱上，并使用富含钾的流动相进行反洗脱，以便通过离子对反相HPLC进行进一步分离。该方法的显著特点包括：（1）将不同的HPLC保留机制在线结合用于儿茶酚胺的富集和分离，以实现最佳的分析特异性；（2）基于1 ml的分析血浆体积，每毫升血浆中去甲肾上腺素和肾上腺素的检测下限低至25 pg；（3）使用向“类似”基质中加入标准品的方法生成校准曲线；（4）由于缺乏晚洗脱的色谱干扰，每日样品通量高；（5）使用优化的氧化铝预纯化程序。讨论了该方法相对于其他用于血浆去甲肾上腺素和肾上腺素的HPLC-EC分析方法的优势，以及在定义分析性能特征时需要考虑的一般因素。
