Analysis of catecholamines and related compounds in one whole metabolic pathway with high performance liquid chromatography based on derivatization

Publication date: November 2019Source: Arabian Journal of Chemistry, Volume 12, Issue 7Author(s): Xiao Huang, Xiao-Feng Guo, Hong Wang, Hua-Shan ZhangAbstractThe simultaneous determination of Tyr, catecholamines and their metabolites in one whole metabolic pathway using high performance liquid chromatography coupled with fluorescence detection (HPLC–FLD) based on pre-column derivatization has been achieved successfully. 1,3,5,7-Tetramethyl-8-(N-hydroxysuccinimidyl butyric ester)-difluoroboradiaza-s-indacene (TMBB-Su), a highly reactive fluorescent reagent synthesized in our previous work, was used for the labeling of tyrosine (Tyr), l-3,4-dihydroxyphenylalanine (l-DOPA), dopamine (DA), norepinephrine (NE), epinephrine (E) and metanephrine (MN). Derivatization conditions including reagent concentration, buffer, reaction temperature and reaction time were also investigated to improve the derivatization efficiency and thus the sensitivity of the detection. The separation of the derivatives was obtained on C18 column with the mobile phase of 20 mM pH 3.5 citric acid (H3Cit)–sodium hydrogen phosphate (Na2HPO4) buffer and methanol. Good linearities with correlation coefficients square (R2) greater than 0.998 in the corresponding concentration ranges were observed and the detection limits (S/N = 3) were found in the range from 0.10 to 0.40 nM (l-DOPA: 1.45 nM). The proposed method has been applied to the detection of catecholamines and related compounds ...
Source: Arabian Journal of Chemistry - Category: Chemistry Source Type: research