title: 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.
authors.affiliation: Department of Pharmaceutical Sciences, St. Jude Children's Research Hospital (SJCRH), Memphis, TN 38105-2794, USA.
authors.forename: K Elaine
authors.initials: KE
authors.lastname: Hubbard
authors.forename: Amy
authors.initials: A
authors.lastname: Wells
authors.forename: Thandranese S
authors.initials: TS
authors.lastname: Owens
authors.forename: Michael
authors.initials: M
authors.lastname: Tagen
authors.forename: Charles H
authors.initials: CH
authors.lastname: Fraga
authors.forename: Clinton F
authors.initials: CF
authors.lastname: Stewart
mesh_terms: D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004298:Dopamine / Q000134:cerebrospinal fluid* / Q000378:metabolism; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D008297:Male; D012701:Serotonin / Q000134:cerebrospinal fluid* / Q000378:metabolism
chemical_list: D012701:Serotonin; D004298:Dopamine