高效液相色谱-荧光检测法测定血浆总同型半胱氨酸浓度

目的 :建立测定血浆总同型半胱氨酸的高效液相色谱 (HPLC) 荧光检测分析方法。方法 :用TCEP将血浆中各种形式的同型半胱氨酸 (Hcy)还原成总同型半胱氨酸 (tHcy) ,以光胺作内标 ,巯基特异性荧光衍生剂SBD F对Hcy ,胱胺等含巯基化合物进行柱前衍生 ,然后行HPLC 荧光检测。结果 :本方法所检测的Hcy浓度在 2 .5～ 16 0 μmol·L- 1 范围具有良好的线性关系 (r =0 .9975 ) ,Hcy最低检测限 0 .6 2 5 μmol·L- 1 ,批内变异小于 5 .5 % ,批间变异小于 9.0 % ,平均回收率 98.87%。结论 :本法简便、灵敏、精确、稳定

High-performance liquid chromatographic assay for plasma total homocysteine

Objective To establish a high performance liquid chromatography (HPLC) assay with an internal standard for determining plasma total homocysteine. Methods The Hcy mixed disulfides or protein bound Hcy in the plasma were reduced with TCEP. Subsequently, the thiol compounds in the samples were derivatized with a thiol specific fluorogenic reagent, SBD F. The derivate was then separated and detected by HPLC with fluorometric detection. Results The detection limit was 0.625 μmol·L -1 ; the rate of recovery was 98.87%; the intra assay coefficient of variation (CV) was less than 5.5%; and the inter assay CV was less than 9.0%. Conclusion The method presented in this paper is simple, sensitive, precise, and stable.