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Determination of homocysteine in plasma by liquid chromatography with fluorescence detection
Author(s) -
Gautier François C.,
Berneron Christophe,
Douce Philippe J.
Publication year - 1999
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/(sici)1099-0801(199905)13:3<239::aid-bmc828>3.0.co;2-8
Subject(s) - chemistry , chromatography , trichloroacetic acid , fluorescence , protein precipitation , detection limit , centrifugation , elution , fluorescence spectroscopy , quantitative analysis (chemistry) , high performance liquid chromatography , physics , quantum mechanics
A liquid chromatographic (LC) method to assay homocysteine in plasma at micro‐molar levels is described. The procedure consists of protein precipitation with trichloroacetic acid and a centrifugation step. The supernatant is then derivatized with a thiol‐specific fluorochromophore and LC assayed with fluorescence detection at 385 nm excitation and 515 nm detection. LC analysis is performed by isocratic elution using an acetate buffer (0.05 M , pH 4) as a mobile phase and an alphaBond column. Recoveries based on a five‐point standard curve calculation ranged from 96 to 112%. The limits of detection and quantification reached were 0.8 and 1.57 µmol/L, respectively. Copyright © 1999 John Wiley & Sons, Ltd.