Ubbink J B, Hayward Vermaak W J, Bissbort S
Department of Chemical Pathology, Faculty of Medicine, University of Pretoria, South Africa.
J Chromatogr. 1991 Apr 19;565(1-2):441-6. doi: 10.1016/0378-4347(91)80407-4.
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.
描述了一种快速等度高效液相色谱（HPLC）法，用于测定人血清中总同型半胱氨酸水平。在进行反相HPLC分析之前，血清硫醇用SBD-F（7-氟苯并-2-恶唑-1,3-二氮杂-4-磺酸盐铵）进行衍生化，SBD-F是一种市售的硫醇特异性荧光探针。SBD-同型半胱氨酸的保留对pH敏感，流动相pH为2.1可确保在6分钟内实现血清硫醇的基线分离。该方法简单、灵敏、可重复（批间变异系数为6.6%），非常适合临床病理实验室常规测定血清同型半胱氨酸水平。
