Related Experiment Videos
[High-performance liquid chromatographic assay for plasma total homocysteine]
Chong-wen Dai1, Guang-sen Zhang, Xian-ping Wu
1Molecular Hematology Laboratory, Second Xiangya Hospital, Central South University, Changsha 410011, China.
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.