Rapid high-performance liquid chromatographic assay for total homocysteine levels in human serum.
A rapid, isocratic high-performance liquid chromatographic (HPLC) method is described for the determination of total homocysteine levels in human serum. Prior to reversed-phase HPLC analysis, the serum thiols were derivatized with SBD-F (ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate), a thiolspecific fluorogenic probe which is commercially available. Retention of SBD-homocysteine was sensitive to pH, and a mobile phase pH of 2.1 ensured baseline separation of serum thiols within 6 min. The method is simple, sensitive, reproducible (between-run coefficient of variation of 6.6%) and very suitable for routine determination of serum homocysteine levels in a clinical pathology laboratory.
Ubbink
J B
JB
Department of Chemical Pathology, Faculty of Medicine, University of Pretoria, South Africa.
Hayward Vermaak
W J
WJ
Bissbort
S
S
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006710:Homocysteine / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration
D006710:Homocysteine