CNRS, UMR5292, Lyon Neuroscience Research Center, CMO Team, Lyon F-69000, France; University of Lyon 1, Villeurbanne F-69000, France.
INSERM, U1028, Lyon Neuroscience Research Center, NeuroDialyTics, Lyon F-69000, France.
J Chromatogr B Analyt Technol Biomed Life Sci. 2014 Mar 1;951-952:52-7. doi: 10.1016/j.jchromb.2014.01.023. Epub 2014 Jan 24.
Electrochemical methods are very often used to detect catecholamine and indolamine neurotransmitters separated by conventional reverse-phase high performance liquid chromatography (HPLC). The present paper presents the development of a chromatographic method to detect monoamines present in low-volume brain dialysis samples using a capillary column filled with sub-2μm particles. Several parameters (repeatability, linearity, accuracy, limit of detection) for this new ultrahigh performance liquid chromatography (UHPLC) method with electrochemical detection were examined after optimization of the analytical conditions. Noradrenaline, adrenaline, serotonin, dopamine and its metabolite 3-methoxytyramine were separated in 1μL of injected sample volume; they were detected above concentrations of 0.5-1nmol/L, with 2.1-9.5% accuracy and intra-assay repeatability equal to or less than 6%. The final method was applied to very low volume dialysates from rat brain containing monoamine traces. The study demonstrates that capillary UHPLC with electrochemical detection is suitable for monitoring dialysate monoamines collected at high sampling rate.
电化学方法常用于检测通过常规反相高效液相色谱 (HPLC) 分离的儿茶酚胺和吲哚胺神经递质。本文介绍了一种使用填充有亚 2μm 颗粒的毛细管柱检测小体积脑透析样品中存在的单胺的色谱方法。在优化分析条件后,对这种新的超高效液相色谱 (UHPLC) 电化学检测方法的几个参数(重复性、线性、准确性、检测限)进行了检验。纳洛酮、肾上腺素、血清素、多巴胺及其代谢产物 3-甲氧基酪胺在 1μL 进样体积中得到分离;它们在 0.5-1nmol/L 以上的浓度下被检测到,具有 2.1-9.5%的准确性和低于或等于 6%的日内重复性。最终方法应用于从小鼠脑中收集的含有单胺痕迹的极低体积透析液。该研究表明,带有电化学检测的毛细管 UHPLC 适合于以高采样率监测收集的透析液单胺。