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Simultaneous determination of catecholamines and their metabolites related to Alzheimer's disease in human urine
Author(s) -
Liu Lili,
Li Qing,
Li Naijing,
Ling Junhong,
Liu Ran,
Wang Yixiang,
Sun Lixin,
Chen Xiao Hui,
Bi Kaishun
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201000799
Subject(s) - urine , metabolite , chromatography , chemistry , pharmacology , medicine , biochemistry
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.