Quantitation of homocysteine in human plasma by capillary electrophoresis and laser-induced fluorescence detection.
Homocysteine (Hcy) represents a branching point between the transsulfuration and transmethylation pathway of methionine. A large increase of plasma concentration of Hcy is observed in patients with inherited hyperhomocysteinemia. A moderated increase (above 10 microM) is also observed in various pathological conditions, such as arterial occlusion, hypertension, hyperlipidemia and chronic renal failure. While amino acids were largely studied using capillary electrophoresis with UV or laser-induced fluorescence detection (LIF), thiol-amino acids were not. In this work we present a new approach for testing homocysteine in human plasma using CE-LIF and fluorescein isothiocyanate. The low fluorescence yield of the fluorescein thiocarbamyl (FTC) thiol-amino acids limits, probably, the sensitivity of the detection to 8 x 10(-10) M (instead of 10(-12) M for FTC-arginine).
Laboratoire de Biochimie INSERM U466, Centre Hospitalier Rangueil, Toulouse, France.
E
E
Caussé
R
R
Terrier
S
S
Champagne
M
M
Nertz
P
P
Valdiguié
R
R
Salvayre
F
F
Couderc
D019075:Electrophoresis, Capillary / Q000379:methods*; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D007834:Lasers; D012015:Reference Standards; D013050:Spectrometry, Fluorescence / Q000379:methods*
D006710:Homocysteine