School of Pharmacy, Shenyang Pharmaceutical University, Shenyang, P. R. China.
J Sep Sci. 2011 May;34(10):1198-204. doi: 10.1002/jssc.201000799. Epub 2011 Apr 1.
A simple and specific high-performance liquid chromatography method coupled with fluorescence detection (HPLC-FL) has been developed for the simultaneous determination of L-3,4-dihydroxyphenylalanine, norepinephrine, dopamine, epinephrine and 3,4-dihydroxyphenylacetic acid in human urine. The samples were derivatized by 1,2-diphenylethylenediamine with isoprenaline as internal standard. The factors affecting the fluorescence yield were investigated, including the reaction and separation conditions. The catecholamine derivatives were separated on a Kromasil C(18) column with methanol and sodium acetate buffer as mobile phase. The limits of detection for all catecholamines ranged from 0.2 to 1.1 ng/mL. The linear ranges were from 2.5 to 200 ng/mL except 3,4-dihydroxyphenylacetic acid from 5 to 200 ng/mL. The intra- and interday RSDs for all catecholamines were 1.0-8.0 and 2.1-14%, respectively. The method was successfully applied to determine the catecholamines in human urine from 14 Alzheimer's disease patients and 14 healthy volunteers. It was concluded that the mean levels of catecholamines in urine of Alzheimer's disease patients were all lower than those in healthy volunteers. The cluster analysis and independent samples T-test were used to distinguish the Alzheimer's disease patients and healthy volunteers.
已开发出一种简单而特异的高效液相色谱法（HPLC-FL），可用于同时测定人尿液中的 L-3,4-二羟苯丙氨酸、去甲肾上腺素、多巴胺、肾上腺素和 3,4-二羟苯乙酸。样品通过 1,2-二苯乙烯二胺与异丙肾上腺素衍生化，以内标物。考察了影响荧光产率的因素，包括反应和分离条件。儿茶酚胺衍生物在 Kromasil C(18)柱上用甲醇和乙酸钠缓冲液作为流动相分离。所有儿茶酚胺的检测限范围为 0.2-1.1ng/mL。线性范围为 2.5-200ng/mL，除 3,4-二羟苯乙酸为 5-200ng/mL。所有儿茶酚胺的日内和日间 RSD 分别为 1.0-8.0%和 2.1-14%。该方法成功应用于测定 14 例阿尔茨海默病患者和 14 名健康志愿者的人尿液中的儿茶酚胺。结论为，阿尔茨海默病患者尿液中儿茶酚胺的平均水平均低于健康志愿者。采用聚类分析和独立样本 T 检验区分阿尔茨海默病患者和健康志愿者。
