Determination of dopamine, serotonin, and their metabolites in pediatric cerebrospinal fluid by isocratic high performance liquid chromatography coupled with electrochemical detection

Abstract

A method to rapidly measure dopamine (DA), dihydroxyindolphenylacetic acid, homovanillic acid, serotonin (5-HT) and 5-hydroxyindoleacetic acid concentrations in cerebrospinal fluid (CSF) has not yet been reported. A rapid, sensitive, and specific HPLC method was therefore developed using electrochemical detection. CSF was mixed with an antioxidant solution prior to freezing to prevent neurotransmitter degradation. Separation of the five analytes was obtained on an ESA MD-150 x 3.2 mm column with a flow rate of 0.37 mL/min and an acetonitrile-aqueous (5 : 95, v/v) mobile phase with 75 mM monobasic sodium phosphate buffer, 0.5 mM EDTA, 0.81 mM sodium octylsulfonate and 5% tetrahydrofuran. The optimal electrical potential settings were: guard cell +325 mV, E1 -100 mV and E2 +300 mV. Within-day and between-day precisions were <10% for all analytes and accuracies ranged from 91.0 to 106.7%. DA, 5-HT, and their metabolites were stable in CSF with antioxidant solution at 4 degrees C for 8 h in the autoinjector. This method was used to measure neurotransmitters in CSF obtained from children enrolled on an institutional medulloblastoma treatment protocol.

Endogenous Opioid Activity in the Anterior Cingulate Cortex Is Required for Relief of Pain

“…The probe was perfused with artificial CSF (in mM: 147.0 NaCl, 2.8 KCl, 1.2 MgCl 2 , and 1.2 CaCl 2 ) at a rate of 2.0 l/min using a syringe, pump drive, and controller (MDN-0250, MD-1001, and MDN-1020, respectively; BASi). After a 75-90 min washout period, two baseline and four experimental fractions (30 min/fraction) were collected into prechilled (4°C) Eppendorf tubes containing 1.5 l of 40ϫ antioxidant solution (6.0 mM L-cysteine, 2.0 mM oxalic acid, and 1.3% glacial acetic acid; Hubbard et al, 2010). In some experiments, one baseline and one experimental fraction (90 min/fraction) were collected.…”

No correlation between serotonin and its metabolite 5-HIAA in the cerebrospinal fluid and [11C]AZ10419369 binding measured with PET in healthy volunteers

“…Due to technical limitations in the measurement of serotonin in the CSF (Anderson et al, 1990), its metabolite 5-hydroxyindoleacetic acid (5-HIAA) has been more frequently used in studies of serotonergic activity, especially in relation to psychiatric disorders (Asberg, 1997). However, with improved quantification methods CSF serotonin has been resurrected as an estimation of extracellular serotonin in the brain (Anderson et al, 2002;Hubbard et al, 2010).…”

“…Five milliliter of CSF was collected and immediately centrifuged at 2000 g for 10 min at room temperature, aliqouted in 1 mL aliquots and frozen at 280 C. All CSF samples were free from contamination with blood as determined with erythrocyte counts. The samples were, then, prepared for analysis with high performance liquid chromatography, with Coulochem III electrochemical detector with a 5011A coulometric analytical cell (Dionex Ultimate 3000 series, ThermoFisher Scientific, Stockholm, Sweden) as previously described (Hubbard et al, 2010;Yang and Beal, 2011). The limit of detection was 0.16 ng/mL for serotonin and 0.23 ng/mL for 5-HIAA.…”

Electrochemical and Hyphenated Electrochemical Detectors in Liquid Chromatography and Flow Injection Systems for Drug Compound Analysis

“…Tables 7.2 and 7.3 show the LC-EC and FIA-EC with their hyphenated EC-MS detection systems used in the methods, respectively. Plasma sample [74] (continued) Cerebrospinal fluid [83] (continued) (continued) [107] (continued) Bovine and human serum samples [118] (continued) 7.5 Hyphenated Techniques for LC and FIA Phyto-pharmaceutical formulations [130] (continued) Plasma and urine samples [144] (continued) …”