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Quantification of luminally released serotonin in rat proximal colon by capillary electrophoresis with laser-induced fluorescence detection
Authors:Sheng-da Qi   Shun-lian Tian   Hong-xi Xu   Joseph J. Y. Sung  Zhao-xiang Bian
Affiliation:(1) School of Chinese Medicine, Hong Kong Baptist University, Hong Kong SAR, China;(2) Department of Chemistry, Lanzhou University, Lanzhou, 730000, China;(3) Hong Kong Jockey Club Institute of Chinese Medicine, Hong Kong SAR, China;(4) Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong SAR, China
Abstract:Serotonin (5-hydroxytryptamine, 5-HT) plays vital roles in regulating gastrointestinal functions. Thus, the detection of 5-HT in the gastrointestinal tract is of great importance for biomedical research, medical diagnosis, and pharmaceutical therapy. This paper presents a simple, sensitive, and fast method for the quantification of luminally released serotonin in the feces and tissues of the rat proximal colon by means of capillary electrophoresis with laser-induced fluorescence detection. 5-Carboxyfluorescein N-succinimidyl ester was used for precolumn derivatization of serotonin. The optimal separation and detection conditions were obtained with an electrophoretic buffer containing 60 mM borate (pH 8.90) and an air-cooled argon-ion laser (excitation at 488 nm, emission at 520 nm). The serotonin concentrations in the feces and tissues of proximal colons were analyzed with this method, and the average values of serotonin in the feces samples were 1.951 ± 0.446 ng/mg (male) and 2.095 ± 0.533 ng/mg (female) and 1.397 ± 0.267 ng/mg in rat proximal colon tissues. The results demonstrate that this method can accurately determine luminally released 5-HT in rats.
Keywords:Serotonin  Capillary electrophoresis  Laser-induced fluorescence
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