de Jong J, Tjaden U R, van t Hof W, van Valkenburg C F
J Chromatogr. 1983 Dec 30;282:443-56. doi: 10.1016/s0021-9673(00)91621-2.
A flexible and efficient system is described for reversed-phase ion-pair partition-chromatographic analysis of serotonin, its precursors tryptophan and 5-hydroxytryptophan, its main metabolite 5-hydroxyindoleacetic acid, and tryptamine. The chromatographic system consists of tri-n-butylphosphate as stationary phase and buffered water-methanol mixtures, containing perchlorate, as mobile phases. Retention can be selectively influenced by means of the pH, the perchlorate concentration, and the methanol content of the mobile phase, as well as the temperature of the phase system. The compounds of interest can be separated within 10 min and no interference from catecholamines and derivatives was observed. Compared with electrochemical detection, fluorometric detection yielded more favourable detection limits and was more selective when supernatants of brain tissue homogenates were directly injected. Both detection systems showed inadequate selectivity if urine samples were directly injected, but 5-hydroxyindoleacetic acid could readily be assayed.
描述了一种灵活高效的系统，用于对血清素、其前体色氨酸和5-羟色氨酸、其主要代谢产物5-羟吲哚乙酸以及色胺进行反相离子对分配色谱分析。该色谱系统由磷酸三正丁酯作为固定相，以及含有高氯酸盐的缓冲水-甲醇混合物作为流动相组成。保留时间可通过流动相的pH值、高氯酸盐浓度、甲醇含量以及相系统的温度进行选择性影响。目标化合物可在10分钟内分离，未观察到来自儿茶酚胺及其衍生物的干扰。与电化学检测相比，荧光检测具有更有利的检测限，并且在直接注入脑组织匀浆上清液时更具选择性。如果直接注入尿液样本，两种检测系统的选择性均不足，但5-羟吲哚乙酸很容易进行测定。
