Independent Optimization of Capillary Electrophoresis Separation and Native Fluorescence Detection Conditions for Indolamine and Catecholamine Measurements

Park, Young Hun; Zhang, Xin; Rubakhin, Stanislav S.; Sweedler, Jonathan V.

doi:10.1021/ac990659r

Cited by 104 publications

(67 citation statements)

References 27 publications

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“…The laboratory-assembled capillary electrophoresis (CE) system with 257 nm excitation and a charge-coupled device/spectrograph for wavelength-resolved fluorescence emission detection was employed for ultra sensitive detection of 5-HT and its metabolites (Fuller et al 1998;Park et al 1999;Zhang et al 2001;Zhang et al 2002;Stuart et al 2003). Briefly, approximately 3 nL of sample was injected onto a 50-lm inner diameter/150-lm outer diameter capillary and separated by an applied voltage of 21-25 kV.…”

Section: Capillary Electrophoresismentioning

confidence: 99%

Systemic serotonin sulfate in opisthobranch mollusks

Stuart¹,

Ebaugh²,

Copes³

et al. 2004

Journal of Neurochemistry

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Serotonin (5-hydroxytryptamine, 5-HT) is a ubiquitous modulatory neurotransmitter with roles as a neurohormone and neurotransmitter. However, few studies have been performed characterizing this molecule and its related metabolites in circulating fluids. Here, we demonstrate native 5-HT sulfate, but much lower levels of 5-HT, in hemolymph of the marine mollusk Pleurobranchaea californica. The metabolite 5-HT sulfate forms from 5-HT uptake and metabolism in central ganglia of Aplysia californica and in the visceral nerve and eye of Pleurobranchaea, but not in hemolymph itself. In addition, 5-hydroxyindole acetic acid (5-HIAA), while not detected in hemolymph, forms in higher quantities than does 5-HT sulfate in the eye and visceral nerve, and c-glu-5-HT is also observed in this area but never in hemolymph. As systemic 5-HT sulfate appears not to originate from the optic region or from systemic 5-HT, 5-HT sulfate likely derives from the nervous system. Circulating 5-HT sulfate is at least 10-fold higher during the light portion of a 12 : 12-h light/dark cycle than during the dark portion (p < 0.0007), but there is no obvious trend for free systemic tryptophan (Trp) (p > 0.3) in Pleurobranchaea. 5-HT in mollusks is associated with general arousal state; thus, diurnal systemic changes in a 5-HT catabolite may reflect a regulatory role for indole catabolism in behavioral rhythms. Keywords: capillary electrophoresis, hemolymph, marine mollusk, serotonin, sulfation. Serotonin (5-hydroxytryptamine, 5-HT) is a biogenic amine responsible for a variety of tasks in the nervous system. For instance, 5-HT is implicated in learning and memory in the central nervous system (CNS) and emesis and peristaltic reflex in the enteric nervous system (ENS) (Bartsch et al. 2000;Buhot et al. 2000;Endo et al. 2000). Several organs serve as targets for 5-HT, but the fate of 5-HT in target tissues, let alone the physiological effects of the resulting metabolites, has not been fully ascertained. Knowledge of the products of transformed 5-HT, as well as their distribution patterns, will assist in illuminating the processes leading to impaired neurotransmission and eventually aid in designing compounds that could potentially inhibit disease in neural and surrounding tissues.Marine mollusks present an excellent model for the study of the nervous system because of their large cell somata and well-characterized neural networks. In gastropod mollusks, 5-HT is involved in many neurophysiological processes, including sensitization and facilitation of withdrawal reflexes (Hawkins et al. 1993), feeding behavior (Gillette and Davis 1977) and locomotion (McPherson and Blankenship 1991). However, 5-HT-related metabolites have received considerably less attention.Our laboratory recently demonstrated that 5-HT metabolism in Aplysia californica involves sulfation and c-glutamylation with trace amounts of monoamine oxidase activity present in the form of 5-hydroxyindole acetic acid (5-HIAA) (Stuart et al. 2003). In particular, 5-HT sulfate is present ...

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Section: Capillary Electrophoresismentioning

confidence: 99%

Systemic serotonin sulfate in opisthobranch mollusks

Stuart¹,

Ebaugh²,

Copes³

et al. 2004

Journal of Neurochemistry

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“…Several detection techniques in CE, including LIF [20][21][22], UV [23,24], MS [25], EC [26][27][28], and fluorescence [29], have been developed to determine catecholamines. Although LIF detection has the advantage of high sensitivity, the disadvantages are expensive instrumentation and complicated approach.…”

Section: Introductionmentioning

confidence: 99%

Determination of catecholamines by CE with direct chemiluminescence detection

Liu

Wang

et al. 2007

Electrophoresis

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A rapid and sensitive method to detect three catecholamines, isoprenaline, epinephrine, and dopamine, by CE coupled with direct luminol-potassium periodate chemiluminescence (CL) detection is described. The conditions for CE separation and CL reaction were systematically optimized. Under the optimum conditions, the baseline separation of three catecholamines was achieved within 6.5 min. The LODs obtained in standard solution were 5.3 6 10 -8 mol/L for isoprenaline, 4.7 6 10 -8 mol/L for epinephrine, and 1.5 6 10 -7 mol/L for dopamine. The RSD of the migration time and peak area were less than 1.8 and 3.6% (n = 5), respectively. The present method was applied to the determination of the dopamine in urine samples of cigarette smokers and nonsmokers. The results obtained indicate that there is a close relationship between the content of dopamine in human urine and the amount of cigarettes smoked daily; the level of dopamine in smokers is higher than in nonsmokers.

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“…A laboratory-assembled CE instrument with LIF detection, as previously described (Fuller et al, 1998;Park et al, 1999;Zhang et al, 2001), was used for this study. Briefly, approximately 3nl of sample was electrokinetically (2.5V) injected onto a capillary with an inner diameter of 50m and an outer diameter of 150m.…”

Section: Materials and Methods Capillary Electrophoresismentioning

confidence: 99%

Serotonin and its metabolism in basal deuterostomes: insights fromStrongylocentrotus purpuratusandXenoturbella bocki

Squires

Rubakhin

Wadhams

et al. 2010

Journal of Experimental Biology

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SUMMARY Serotonin (5-HT), an important molecule in metazoans, is involved in a range of biological processes including neurotransmission and neuromodulation. Both its creation and release are tightly regulated, as is its removal. Multiple neurochemical pathways are responsible for the catabolism of 5-HT and are phyla specific; therefore, by elucidating these catabolic pathways we glean greater understanding of the relationships and origins of various transmitter systems. Here, 5-HT catabolic pathways were studied in Strongylocentrotus purpuratus and Xenoturbella bocki, two organisms occupying distinct positions in deuterostomes. The 5-HTrelated compounds detected in these organisms were compared with those reported in other phyla. In S. purpuratus, 5-HT-related metabolites include N-acetyl serotonin, -glutamyl-serotonin and 5-hydroxyindole acetic acid; the quantity and type were found to vary based on the specific tissues analyzed. In addition to these compounds, varying levels of tryptamine were also seen. Upon addition of a 5-HT precursor and a monoamine oxidase inhibitor, 5-HT itself was detected. In similar experiments using X. bocki tissues, the 5-HT-related compounds found included 5-HT sulfate, -glutamyl-serotonin and 5-hydroxyindole acetic acid, as well as 5-HT and tryptamine. The sea urchin metabolizes 5-HT in a manner similar to both gastropod mollusks, as evidenced by the detection of -glutamyl-serotonin, and vertebrates, as indicated by the presence of 5-hydroxyindole acetic acid and N-acetyl serotonin. In contrast, 5-HT metabolism in X. bocki appears more similar to common protostome 5-HT catabolic pathways.

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Independent Optimization of Capillary Electrophoresis Separation and Native Fluorescence Detection Conditions for Indolamine and Catecholamine Measurements

Cited by 104 publications

References 27 publications

Systemic serotonin sulfate in opisthobranch mollusks

Systemic serotonin sulfate in opisthobranch mollusks

Determination of catecholamines by CE with direct chemiluminescence detection

Serotonin and its metabolism in basal deuterostomes: insights fromStrongylocentrotus purpuratusandXenoturbella bocki

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