de Jong J, Tjaden U R, Visser E, Meijer W H
Gorlaeus Laboratories, University of Leiden, The Netherlands.
J Chromatogr. 1987 Aug 7;419:85-94. doi: 10.1016/0378-4347(87)80268-2.
The determination of serotonin and 5-hydroxyindoleacetic acid in urine is described. The compounds are separated by means of reversed-phase chromatography using tri-n-butylphosphate as the stationary phase and a mobile phase containing perchlorate as an ion-pairing agent. Detection is performed fluorimetrically. The compounds can be assayed directly, using two mobile phases with different pH values. Alternatively, both compounds can be determined with the same mobile phase after urine extraction on a small XAD-2 column for clean-up of serotonin. Two different approaches were evaluated for this XAD-2 extraction, viz.: (i) desorption of serotonin with a methanol-rich solvent after a washing step with an aqueous solvent; (ii) direct desorption of serotonin in the washing step using a solvent with a small percentage of methanol. Only the latter approach proved to be successful. The linearity of the method was investigated by standard additions to urine. The method was applied to the determination of both compounds in urine. The stability of the compounds in standard solutions and in urine was investigated under various storage conditions.
本文描述了尿液中血清素和5-羟吲哚乙酸的测定方法。采用反相色谱法，以磷酸三正丁酯为固定相，含高氯酸盐作为离子对试剂的流动相进行化合物分离。采用荧光法进行检测。化合物可直接测定，使用两种不同pH值的流动相。或者，在尿液通过小型XAD - 2柱萃取以净化血清素后，两种化合物可用相同的流动相进行测定。针对这种XAD - 2萃取评估了两种不同方法，即：(i) 在用水性溶剂洗涤步骤后，用富含甲醇的溶剂解吸血清素；(ii) 在洗涤步骤中使用含少量甲醇的溶剂直接解吸血清素。仅后一种方法被证明是成功的。通过向尿液中加入标准品研究了该方法的线性。该方法应用于尿液中两种化合物的测定。在各种储存条件下研究了化合物在标准溶液和尿液中的稳定性。
