Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Anal Bioanal Chem. 2012 Jan;402(3):1393-7. doi: 10.1007/s00216-011-5542-x. Epub 2011 Nov 11.
A liquid chromatography-chemiluminescence detection method was developed and validated for the determination of catecholamines (norepinephrine, epinephrine, and dopamine) in mouse brains. Chromatography was performed on a strong cation exchange column (150 × 2.0-mm id) using an isocratic mobile phase of 65 mM potassium acetate/75 mM potassium phosphate (95:5, pH 3.5) at a flow rate of 0.2 mL/min following post-column fluorescence derivatization of catecholamines with ethylenediamine and peroxyoxalate chemiluminescence reaction detection. The recovery of catecholamines added to mouse brain samples was more than 95.0%, while intra- and inter-day precision of the assay were <4.8%. The validated method was used to determine norepinephrine and dopamine concentrations in mouse brains without prior sample purification.
建立并验证了一种用于测定小鼠脑内儿茶酚胺（去甲肾上腺素、肾上腺素和多巴胺）的高效液相色谱-化学发光检测法。色谱分析采用强阳离子交换柱（150×2.0-mm id），以 65mM 乙酸钾/75mM 磷酸钾（95:5，pH3.5）为等度流动相，流速为 0.2mL/min，在柱后与乙二胺衍生化，并通过过氧草酸化学发光反应检测儿茶酚胺。加入到小鼠脑样中的儿茶酚胺的回收率大于 95.0%，而该测定法的日内和日间精密度均<4.8%。该方法无需样品纯化即可直接测定小鼠脑内去甲肾上腺素和多巴胺的浓度。
