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A sensitive and selective determination method of histamine by HPLC with intramolecular excimer‐forming derivatization and fluorescence detection
Author(s) -
Yoshitake Takashi,
Ichinose Fumio,
Yoshida Hideyuki,
Todoroki Kenichiro,
Kehr Jan,
Inoue Osamu,
Nohta Hitoshi,
Yamaguchi Masatoshi
Publication year - 2003
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.264
Subject(s) - chemistry , derivatization , chromatography , high performance liquid chromatography , acetonitrile , triethylamine , detection limit , reagent , fluorescence , histamine , organic chemistry , medicine , physics , quantum mechanics
A highly sensitive, selective and simple method is described for the determination of histamine by high‐performance liquid chromatography (HPLC) with uorescence detection. The method is based on an intramolecular excimer‐forming uorescence derivatization of histamine with 4‐(1‐pyrene)butyric acid N ‐hydroxysuccinimide ester (PSE), followed by reversed‐phase HPLC. Histamine, having two amino moieties in a molecule, was converted to the dipyrene‐labeled derivative by reaction with PSE. The derivative afforded intramolecular excimer uorescence (450–540 nm), which can clearly be discriminated from the monomer uorescence (370–420 nm) emitted from PSE. Typically, a 10 µL sample solution was mixed with 100 µL of derivatization reagent solution, which was a mixture of 0.5 m m PSE in acetonitrile and 0.5 m m potassium carbonate in water (8:2, v/v). The derivatization was carried out at 100°C for 90 min. The PSE derivative of histamine could be separated by reversed‐phase ODS column with isocratic elution using acetonitrile:water (82:18, v/v) containing 0.03% triethylamine. The detection limit (singnal‐to‐noise ratio = 3) of histamine was 0.5 fmol for a 30 µL injection. The method was successfully applied to the determination of histamine in human urine, and had enough selectivity and sensitivity for urinary histamine quantication. Copyright © 2003 John Wiley & Sons, Ltd.