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A Modified Method For The Determination Of Plasma Total Homocysteine By High Performance Liquid Chromatography, Serhat Tokgöz, Cumhur Bi̇lgi̇, Erdi̇nç Çakir, M. Kemal Erbi̇l, Türker Kutluay
A Modified Method For The Determination Of Plasma Total Homocysteine By High Performance Liquid Chromatography, Serhat Tokgöz, Cumhur Bi̇lgi̇, Erdi̇nç Çakir, M. Kemal Erbi̇l, Türker Kutluay
Turkish Journal of Medical Sciences
We have modified a high-performance liquid chromatography (HPLC) procedure for homocysteine assay using 7-fluoro-2, 1, 3-benzoxadiazole-4-sulfonamide (ABD-F) pre-column derivatization. In this study, the thiol compounds were liberated from plasma proteins by reduction with 2-mercaptoethanol (BME) and derivatized with a thiol-specific fluoregenic marker, ABD-F. The derivatives were separated by reversed-phase HPLC on a Hypersil C 18 column (4.6 x 150 mm; 5 µ) using isocratic elution with 0.1 M phosphate buffer pH 6.0 containing 8% methanol and fluorescence detection. Excitation and emission wavelengths were 386 and 516 nm. The peak of homocysteine was eluted at 413 ± 25 s. Mean within-day …