Chromatographic analysis of serotonin, 5-hydroxyindolacetic acid and homovanillic acid in dried blood spots and platelet poor and rich plasma samples

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• 作者：Maria Addolorata Saracino ; Gilberto Gerra ; Lorenzo Somaini ; Marco Colombati ; Maria Augusta Raggi
• 关键词：Serotonin ; 5-Hydroxyindolacetic acid ; Homovanillic acid ; High-performance liquid chromatography ; Dried blood spots ; Platelet poor and rich plasma
• 刊名：Journal of Chromatography A
• 出版年：2010
• 出版时间：16 July 2010
• 年：2010
• 卷：1217
• 期：29
• 页码：4808-4814
• 全文大小：430 K

文摘

An isocratic high-performance liquid chromatographic method has been developed for the measurement of serotonin, 5-hydroxyindolacetic and homovanillic acids in dried blood spots and in platelet poor and rich plasma samples. Analyses were carried out on a C18 reversed-phase column using a mobile phase composed of 13 % methanol and 87 % aqueous citrate buffer, containing octanesulfonic and ethylendiaminotetracetic acids. Coulometric detection was used, setting the guard cell at +0.100 V, the first analytical cell at −0.200 V and the second analytical cell at +0.400 V. For the pre-treatment of biological samples a novel solid-phase extraction procedure, based on mixed-mode reversed-phase – strong anion exchange Oasis cartridges, was implemented. Extraction yields of the analytes from all these matrices were satisfactory, being always higher than 89.0 % . The calibration curve was linear over the on-column concentration range of 0.1–22.5 ng mL⁻¹ for serotonin and 5-hydroxyindolacetic acid and of 0.25–22.5 ng mL⁻¹ for homovanillic acid. The sensitivity was good with a limit of detection of 0.05 ng mL⁻¹ for serotonin and 5-hydroxyindolacetic acid and 0.12 ng mL⁻¹ for homovanillic acid. Results were also satisfactory in terms of precision, selectivity and accuracy. The analytical method was successfully applied to human platelet poor and rich plasma samples and to dried blood spots from volunteers and psychiatric patients.