title: Development of analytical method for catechol compounds in mouse urine using hydrophilic interaction liquid chromatography with fluorescence detection.
abstract: An analytical method for catecholamines and related compounds using hydrophilic interaction liquid chromatography (HILIC) with native fluorescence detection has been developed. We found that ZIC-cHILIC with phosphorylcholine was suitable for the separation of catechol compounds with good peak shapes among six different HILIC columns (Inertsil SIL, Inertsil Amide, Inertsil Diol, TSKgel NH2-100, ZIC-HILIC, and ZIC-cHILIC). Using ZIC-cHILIC, eight catechol compounds (dopamine, epinephrine, norepinephrine, 3,4-dihydroxyphenylalanine, 3,4-dihydroxyphenylacetic acid, 3,4-dihydroxyphenylglycol, 3,4-dihydroxymandelic acid, and internal standard 3,4-dihydroxybenzylamine) were separated within 15min. The limit of detection at a signal to noise ratio of 3 was 3-28nM. An improved sensitivity was obtained as compared to that of reversed-phase liquid chromatography. This was partly attributed to the increase in the fluorescence intensity of the catechol compounds in the acetonitrile-rich mobile phase. Solid phase extraction using a monolithic silica disk-packed spin column with phenylboronate moieties, which have affinity to catechol compounds, was performed for the selective extraction of catechol compounds from mouse urine. Dopamine, epinephrine, norepinephrine, 3,4-dihydroxyphenylalanine, and 3,4-dihydroxyphenylglycol were successfully quantified in mouse urine.
authors: Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan.
authors: Takahiro
authors: T
authors: Kanamori
authors: Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan.
authors: Muneki
authors: M
authors: Isokawa
authors: Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan.
authors: Takashi
authors: T
authors: Funatsu
authors: Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan. Electronic address: makotot@mol.f.u-tokyo.ac.jp.
authors: Makoto
authors: M
authors: Tsunoda
mesh_terms: D000818:Animals; D002396:Catechols / Q000652:urine*; D002853:Chromatography, Liquid / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D002396:Catechols; C034221:catechol
keywords: Catecholamines; Monolithic silica; Phenylboronate; ZIC-cHILIC