Hubbard K Elaine, Wells Amy, Owens Thandranese S, Tagen Michael, Fraga Charles H, Stewart Clinton F
Department of Pharmaceutical Sciences, St. Jude Children's Research Hospital (SJCRH), Memphis, TN 38105-2794, USA.
Biomed Chromatogr. 2010 Jun;24(6):626-31. doi: 10.1002/bmc.1338.
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.
尚未报道一种快速测量脑脊液（CSF）中多巴胺（DA）、二羟基吲哚苯乙酸、高香草酸、5-羟色胺（5-HT）和5-羟基吲哚乙酸浓度的方法。因此，我们开发了一种使用电化学检测的快速、灵敏且特异的高效液相色谱（HPLC）方法。在冷冻前将脑脊液与抗氧化剂溶液混合，以防止神经递质降解。在一根ESA MD-150×3.2 mm柱上实现了5种分析物的分离，流速为0.37 mL/min，流动相为乙腈-水（5:95，v/v），含有75 mM磷酸二氢钠缓冲液、0.5 mM乙二胺四乙酸（EDTA）、0.81 mM辛烷磺酸钠和5%四氢呋喃。最佳电势设置为：保护电极+325 mV，E1 -100 mV，E2 +300 mV。所有分析物的日内和日间精密度均<10%，准确度范围为91.0%至106.7%。DA、5-HT及其代谢产物在含有抗氧化剂溶液的脑脊液中于4℃在自动进样器中可稳定8小时。该方法用于测量从参加机构性髓母细胞瘤治疗方案的儿童中获取的脑脊液中的神经递质。
