血浆总同型半胱氨酸的高效液相色谱测定法
[High-performance liquid chromatographic assay for plasma total homocysteine].
作者信息
Dai Chong-wen, Zhang Guang-sen, Wu Xian-ping
机构信息
Molecular Hematology Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China.
出版信息
Hunan Yi Ke Da Xue Xue Bao. 2002 Dec 28;27(6):556-8.
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 mumol.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.
目的
建立一种采用内标法的高效液相色谱（HPLC）法测定血浆总同型半胱氨酸。
方法
血浆中的同型半胱氨酸混合二硫化物或与蛋白质结合的同型半胱氨酸用三（2-羧乙基）膦（TCEP）还原。随后，样品中的硫醇化合物用硫醇特异性荧光试剂SBD-F进行衍生化。然后通过高效液相色谱荧光检测法对衍生物进行分离和检测。
结果
检测限为0.625μmol·L-1；回收率为98.87%；批内变异系数（CV）小于5.5%；批间CV小于9.0%。
结论
本文提出的方法简单、灵敏、精确且稳定。
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